人β防御素3和植物甜蛋白des-pGlu1-Brazzein基因的重组表达研究
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摘要
食品安全一直是人们关注的焦点,安全、天然的食品添加剂是保障食品安全的重要方面。人β防御素3(hBD_3)因其广谱高效的杀菌作用和较低的细胞毒性,有望成为新型食品防腐剂。植物des-pGlul-Brazzein甜蛋白因其高甜度和对pH、热的稳定性而有望成为新型甜味剂。本课题以大肠杆菌和甲醇酵母为宿主重组表达了hBD_3、des-pGlul-Brazzein以及两者的嵌合基因,并采用摇瓶发酵分析了适合三株重组大肠杆菌菌株生长和目的蛋白表达的发酵条件。主要结果如下:
采用细菌和酵母偏爱密码子分别合成了hBD_3、des-pGlul-Brazzein及两者的嵌合基因片段(中间加了一段编码凝血酶酶切位点的片段),这些编码序列被插入pET30a和pPIC9K载体,分别构建成重组载体pET-hBD_3、pET-Bra、pET-hBD_3-Bra和pPIC9K-hBD_3、pPIC9K-Bra、pPIC9K-hBD_3-Bra。
以E.coli为重组表达宿主,pET30a为表达载体,对hBD_3进行了融合表达,重组hBD_3融合蛋白占总蛋白的30.9%,实现了目的蛋白的高效表达。通过亲和层析和进一步酶切纯化得到了较纯的hBD_3蛋白,无论是重组hBD_3融合蛋白还是hBD_3蛋白对革兰氏阳性菌金黄色葡萄球菌和革兰氏阴性菌大肠杆菌都具有较高的抑菌活性,并且酶切后得到的hBD_3蛋白的活性比重组hBD_3融合蛋白高,完全可以与提取得到的天然hBD_3的活性相媲美。
以E.coli为重组表达宿主,pET30a为表达载体,对des-pGlul-Brazzein进行了融合表达,重组des-pGlul-Brazzein融合蛋白占总蛋白的35%以上,通过亲和层析和进一步酶切纯化得到了较纯的des-pGlul-Brazzein蛋白,两者的相对甜度分别是蔗糖的400和600倍,在80℃作用4h后其甜味并未丧失,说明得到的甜蛋白具有较强热稳定性。
以E.coli为重组表达宿主,pET30a为表达载体,对嵌合基因hBb_3-Bra进行了融合表达。hBD_3-Bra融合蛋白占总蛋白的30.5%,实现了在大肠杆菌中的高效表达。融合蛋白只有很弱的杀菌活性,但甜度约为蔗糖200倍,通过凝血酶切割纯化后,得到了具有明显杀菌活性的重组hBD_3融合蛋白和甜度约为蔗糖600倍的des-pGlul-Brazzein蛋白。
Food safety is one of the hot spot all the time, and the natural & safe food additive is the key to assuring food safety. Human p-defensin-3 (hBD_3) is expected to be developed as a new food antimicrobial additive because of its broad-spectrum antimicrobial activity and low cytotoxicity to human. The sweet-tasting protein des-pGlul-Brazzein, isolated from the fruit of Pentadiplandra brazzeana Baillion, is expected to be developed as a new food sweet additive because of its high sweetness and heat & pH-stability. The project aimed at: (1) High-level expression of hBD_3, des-pGlul-Brazzein and hBD_3-Bra mosaic gene in E.coli and Pichia pastoris respectively; (2) Optimizing the fermentation conditions of the target strains by flask shake. The main results are shown as following:According to Escherichia and yeast biased codon, the coding sequence of hBD_3, des-pGlul-Brazzein and mosaic gene hBD_3-Bra ( connected with the sequence coding thrombin cleavage site) were constructed respectively, and were inserted into the multiclone site in pET30a and pPIC9K vector respectively, resulting in the recombinant expression vectors pET-hBD_3, pEl-Bra, pET-hBD_3-Bra and pPIC9K-hBD_3, pPlC9K-Bra, pPIC9K-hBD_3-Bra.The fusion protein of hBD_3 was expressed by using pET30a as expression vector and E.coli as host, the recombinant protein was expressed after induction by IPTG, which accounted for 30.9% of total protein of host cell. The recombinant fusion protein of hBD3 was purified by affinity column and then digested by enterokinase and purified by affinity column again, resulting in the recombinant natural hBD_3. The two interested protein both have high antimicrobial activity against E.coli and S.aureus, similar as natural protein.The fusion protein of des-pGlul-Brazzein was expressed by using pET30a as expression vector and E.coli as host, the recombinant protein was expressed after induction by IPTG, which accounted for 35% of total protein of host cell. The recombinant fusion protein of des-pGlul-Brazzein was purified by affinity column and then digested by enterokinase and purified by affinity column again, resulting in the recombinant natural des-pGlul-Brazzein. The sweetness of the two interested protein is 200 times and 600 times than that of sucrose respectively, including kept after 4h at 80 °C.
The chimeric protein of hBD3-Bra was expressed by using pET30a as expression vector and E.coli as host, the recombinant chimeric protein of hBD3-Bra was expressed after induction by IPTG, which accounted for 30.5% of total protein of host cell. hBD3-Bra has a weak antimicrobial activity, but is 200 times sweeter than that of sucrose. After digested by thrombin and purified by affinity column, the recombinant hBD3 has a high antimicrobial activity, and the natural des-pGlul-Brazzein also has 600-time sweetness of sucrose, much sweeter than that of fusion protein Most of the expressed proteins were in inclusion bodies, some was soluble. Both the soluble and renaturation protein from inclusion bodies have their own activity whether by dialysis or by column.The fermentation conditions for BL21-pET-hBD_3, BL21-pET-Bra and BL2l-pET-hBD3-Bra were optimized respectively by flask shake. The best conditions for growth and fermentation were basically same, i.e. culture volume 100mL (500mL flask), seed dose 1.5%, medium original pH value 7.0. The culture volume was the most important factor. The growth speed would be lower as the culture volume increased more. It suggested the content of soluble O2 is key factor for growth and fermentation of the strains,. The biomass would increase as time passed.In order to study the influence of different concentration of inducer, temperature and time on the growth and the yield of target protein of strain respectively, the strains were induced by IPTG and lactose at 37°C and 30°C for 4h and 6h respectively. The results showed that the concentration of IPTG had no obvious effect on the growth and the yield of expression of strains between 0.2-lmM, The concentration of lactose obviously inhibit the growth of s
采用细菌和酵母偏爱密码子分别合成了hBD_3、des-pGlul-Brazzein及两者的嵌合基因片段(中间加了一段编码凝血酶酶切位点的片段),这些编码序列被插入pET30a和pPIC9K载体,分别构建成重组载体pET-hBD_3、pET-Bra、pET-hBD_3-Bra和pPIC9K-hBD_3、pPIC9K-Bra、pPIC9K-hBD_3-Bra。
以E.coli为重组表达宿主,pET30a为表达载体,对hBD_3进行了融合表达,重组hBD_3融合蛋白占总蛋白的30.9%,实现了目的蛋白的高效表达。通过亲和层析和进一步酶切纯化得到了较纯的hBD_3蛋白,无论是重组hBD_3融合蛋白还是hBD_3蛋白对革兰氏阳性菌金黄色葡萄球菌和革兰氏阴性菌大肠杆菌都具有较高的抑菌活性,并且酶切后得到的hBD_3蛋白的活性比重组hBD_3融合蛋白高,完全可以与提取得到的天然hBD_3的活性相媲美。
以E.coli为重组表达宿主,pET30a为表达载体,对des-pGlul-Brazzein进行了融合表达,重组des-pGlul-Brazzein融合蛋白占总蛋白的35%以上,通过亲和层析和进一步酶切纯化得到了较纯的des-pGlul-Brazzein蛋白,两者的相对甜度分别是蔗糖的400和600倍,在80℃作用4h后其甜味并未丧失,说明得到的甜蛋白具有较强热稳定性。
以E.coli为重组表达宿主,pET30a为表达载体,对嵌合基因hBb_3-Bra进行了融合表达。hBD_3-Bra融合蛋白占总蛋白的30.5%,实现了在大肠杆菌中的高效表达。融合蛋白只有很弱的杀菌活性,但甜度约为蔗糖200倍,通过凝血酶切割纯化后,得到了具有明显杀菌活性的重组hBD_3融合蛋白和甜度约为蔗糖600倍的des-pGlul-Brazzein蛋白。
Food safety is one of the hot spot all the time, and the natural & safe food additive is the key to assuring food safety. Human p-defensin-3 (hBD_3) is expected to be developed as a new food antimicrobial additive because of its broad-spectrum antimicrobial activity and low cytotoxicity to human. The sweet-tasting protein des-pGlul-Brazzein, isolated from the fruit of Pentadiplandra brazzeana Baillion, is expected to be developed as a new food sweet additive because of its high sweetness and heat & pH-stability. The project aimed at: (1) High-level expression of hBD_3, des-pGlul-Brazzein and hBD_3-Bra mosaic gene in E.coli and Pichia pastoris respectively; (2) Optimizing the fermentation conditions of the target strains by flask shake. The main results are shown as following:According to Escherichia and yeast biased codon, the coding sequence of hBD_3, des-pGlul-Brazzein and mosaic gene hBD_3-Bra ( connected with the sequence coding thrombin cleavage site) were constructed respectively, and were inserted into the multiclone site in pET30a and pPIC9K vector respectively, resulting in the recombinant expression vectors pET-hBD_3, pEl-Bra, pET-hBD_3-Bra and pPIC9K-hBD_3, pPlC9K-Bra, pPIC9K-hBD_3-Bra.The fusion protein of hBD_3 was expressed by using pET30a as expression vector and E.coli as host, the recombinant protein was expressed after induction by IPTG, which accounted for 30.9% of total protein of host cell. The recombinant fusion protein of hBD3 was purified by affinity column and then digested by enterokinase and purified by affinity column again, resulting in the recombinant natural hBD_3. The two interested protein both have high antimicrobial activity against E.coli and S.aureus, similar as natural protein.The fusion protein of des-pGlul-Brazzein was expressed by using pET30a as expression vector and E.coli as host, the recombinant protein was expressed after induction by IPTG, which accounted for 35% of total protein of host cell. The recombinant fusion protein of des-pGlul-Brazzein was purified by affinity column and then digested by enterokinase and purified by affinity column again, resulting in the recombinant natural des-pGlul-Brazzein. The sweetness of the two interested protein is 200 times and 600 times than that of sucrose respectively, including kept after 4h at 80 °C.
The chimeric protein of hBD3-Bra was expressed by using pET30a as expression vector and E.coli as host, the recombinant chimeric protein of hBD3-Bra was expressed after induction by IPTG, which accounted for 30.5% of total protein of host cell. hBD3-Bra has a weak antimicrobial activity, but is 200 times sweeter than that of sucrose. After digested by thrombin and purified by affinity column, the recombinant hBD3 has a high antimicrobial activity, and the natural des-pGlul-Brazzein also has 600-time sweetness of sucrose, much sweeter than that of fusion protein Most of the expressed proteins were in inclusion bodies, some was soluble. Both the soluble and renaturation protein from inclusion bodies have their own activity whether by dialysis or by column.The fermentation conditions for BL21-pET-hBD_3, BL21-pET-Bra and BL2l-pET-hBD3-Bra were optimized respectively by flask shake. The best conditions for growth and fermentation were basically same, i.e. culture volume 100mL (500mL flask), seed dose 1.5%, medium original pH value 7.0. The culture volume was the most important factor. The growth speed would be lower as the culture volume increased more. It suggested the content of soluble O2 is key factor for growth and fermentation of the strains,. The biomass would increase as time passed.In order to study the influence of different concentration of inducer, temperature and time on the growth and the yield of target protein of strain respectively, the strains were induced by IPTG and lactose at 37°C and 30°C for 4h and 6h respectively. The results showed that the concentration of IPTG had no obvious effect on the growth and the yield of expression of strains between 0.2-lmM, The concentration of lactose obviously inhibit the growth of s
引文
1. Abe K, Yamashita H, Arai S, Kurihara Y. Molecular cloning of curculin, a novel taste-modifying protein with a sweet taste. Biochem Biophys Acta. 1992, 1130: 232-234
1. Andersson SGE, Kurland CG. Coden preferences in free-living microorganisms. Microbiological review. 1990(6), 198-210
2. Assadi-porte FM, Aceti DJ, Cheng H, Markley J L. Efficient production of recombinant Brazzein, a small, heat-stable, sweet-tasting protein of plant origin. Archives of Biochemistry Biophysics. 2000, 376(2): 252-258
3. Assadi-porter F M, Aceti D J, Markley J L. Sweetness determinant sites of Brazzein, a small, heat-stable, sweet-tasting protein. Archives of Biochemistry Biophysics. 2000, 376(2): 259-265
4. Antcheva N, Bonitto M, Zelezetsky I, et al. Effects of positively selected sequence variations in human and Macaca fascicularis β defensin 2 on antimicrobial activity. Antimicrob Agents Chemother. 2004, 2: 685-688
5. Bals R, Wang X, Wu Z, et al. Human β-defensin-2 is a salt-sensitive peptide antibiotic expressed in human lung. Journal of clinical investigation. 1998, 102: 874-880
6. Befus A D, Mowat C, Gilchrist M, et al. Neutrophil defensins induce histamine secretion from mast cells: mechanisms of action. J Immunol. 1999, 163: 947-953
7. Bensch KW, Raida M, Magert H J. hBD-1: a novel beta-defensin from human plasma. FEBS Letter. 1995, 368: 331
8. Bevins CL, Jones DE, Dutra A, Schaffzin J, Muenke M. Human enteric defensin genes: chromosome map position and a model for possible evolutionary relationships. Genomics. 1996, 31: 95-106
9. Boniotto M, Antcheva N, Zelezetsky I, Tossi A, Palumbo V, et al. A study of host defence peptide β-defensin 3 in primates. Biochemisty. 2003, 374: 707-714
10. Boniotto M, Tossi A, Del P, Sgubin, et al. Evolution of the β-defensin 2 gene in primates. Genes Immunity. 2003, 4: 251-257
11. Brierley, RA. Secretion of recombinant human insulin-like growth factor I (IGF-1). Methods Mol Biol. 1998, 103: 149-177
12. Bujacz G. Structure of monellin refined to 2.3 a resolution in the orthorhombic crystal form. Acta Crystallogr D Biol Crystallogr. 1997, 53(Pt 6): 713-9.
13.Cai S J(蔡绍晖),Du J(杜军),Chen X N(陈新年)et al.Recombinant human β-denfensin-2 gene expression in transfected COS-7 cell. Journal of West China University of Medical Sciences(华西医大学报).2000,31(4):433—437
14.Cai S J(蔡绍晖),Du J(杜军),Chen X N(陈新年),et al.Expression of recombinant human β-defensin-2 gene with C terminal of double Marks of Myc and poly-histidine in transfected COS-7 cells(C端带Myc和多聚组氨酸双标记的重组人β-防御素-2在COS-7细胞的转染表达).J Biomed Eng(生物医学工程杂志).2003,20(2):255-259
15.Cai S J(蔡绍晖),Huang N(黄宁),Wang B Y(王伯瑶).重组人β-防御素-2基因在昆虫细胞中的转染表达.Chinese joumal of pathophysiology(中国病理生理杂志).2001,17(8):800
16.Cai SH H(蔡绍晖),et al.Construction of plant expressive vector of human β-defensin-2 gene(重组人β-防御素-2基因的植物表达载体的建立).J Sichuan Univ(Med Sci Edi)(四川大学学报(医学版).2003.34(3):385-389
17. Caldwell JE, Abildgaard F, Dzakula Z, et al. Solution structure of the thermostable-sweet-tasting protein brazzein. Nat Stru Biol. 1998, 5-6: 427-431
18. Callewaert N, Larey W, Cadirgi H, et al. Use of HDEL-tagged trichoderma reesei mannosyl oligosaceharide 1,2-alpha-D-mannosidase for N-glycan engineering in Pichia Pastoris. FEBS letter. 2001, 503: 173-178
19. Cereghino Joan Lin, Cregg James M. Heterologous protein express in the methylotrophic yeast Pichia Pastoris. FEMS Microbiology. 2000, 24, 45-66
20. Chan B, Park H S, Kim SC. Mechanism of adion of antimicrobial peptide buforin Ⅱ: buforin Ⅱ kills microorganism by penetrating the all membrance and inhibiting celluar functions. Biochemical and biophysical Research Commucation. 1998, 244: 253-257
21.Chen shan(陈姗),He Fengtian(何凤田),et al.The cloning, high level expression in E. coli of human beta defensin 3 and its antimicrobial activity analysis. Chinese Journal of biotechnology(生物工程学报).2004,20(4):490—495
22.Chen Y(陈颖),Ge Y Q(葛毅强),Zhang L M(张利明),et al.Adyances in the research and Applicatin of mammal defensin,(哺乳动物防御素的研究进展及其应用前景).Prog Biochem Biophys(生物化学与生物物理进展).2001,28(1):17—21
23. Chertov O, Miciel DF, Xu L, Wang JM, et al. Identification of defensin-1, defensin-2, and CAP37/azurocidin as T-cell chemoattractant proteins released from interleukin-8-stimulated neutrophiis. J Biol Chem. 1996, 271, 2935-2940.
24.Chen Yumei(陈玉梅).酿酒酵母工程菌研究进展.生物工程进展.1993,13(1),11-16
25. Chertov O, Michiel D F, Xu L, et al. Identification of defensin-1,defensin-2 and CAP37/azurocidin as T cell chemoattractant proteins released from interleukin-8-stimulated neutrophils. J Biol Chem. 1996, 271: 2935-2940
26. Chung WO, Dale BA. Innate immune response of oral and foreskin keratinoeytes: utilization of different signaling pathways by various bacterial specirs. Infection and immunity. 2004, 1: 352-358
27. Cipalova I, Hostinova E, Gasperik J, Velebny V. High-level espression and purification of recombinant hBD-1 fused to LMN protein in E.coli. Protein Expr Purif. 2004, 37 (1): 207-212
28. Clare J J, Romanos M A, Rayment F B, et al. Production of mouse epidermal growth factor in yeast: high-level secretion using pichia pastries strains containing multiple gene copies. Gene. 1991, 105: 205-212
29. Clare JJ, Rayment FB, Ballantine SP, Screekrishna K, Romanos MA. High-level expression of tetanus toxin fragment C in pichia pastries strains containing multiple tandem integrations of the gene. Biotechnology. 1991, 9: 455-460
30.Cui H-ZH(崔洪志),Li M(李敏),Xu Q-F(徐琼芳),Guo S-D(郭三堆).Synthesis of a Bacterial-like Plant Monellin Gene and its Exptession in E. coli, Scientia Agriculture Sinica (中国农业科学).1999,32(1):58-62
31. Cociancich S, Ghazi A, Hetru C, Hoffmann J A, et al. Insect defensin, an inducible antibacterial peptide, forms voltage-dependent channels in Micrococcus luteus. J Biol Chem. 1993, 268: 19239-19245
32. Cole AM, Dewan P, Ganz T. Innate antimicrobial activity of nasal secrections. Infect Immun. 1999, 67: 3267-3275
33. Cregg JM, Madden KR, Barringer KJ. Function characterization of the two alcohol oxidase genes from the yeast Pichia Pastoris. Mol Cell Biol. 1989, 9: 1316-1323
34. Cregg JM, Vedvick T, Raschke W C, Recent advance in the expression of foreign genes in Pichia Pastoris. Biotechnology. 1993, 11: 905-910
35. Daniell S, Mellits K H, et al. Refolding the sweet-tasting protein thaumatin Ⅱ from insoluble bodies synthesized in E.coli. Food Chemistry. 2000, 71: 105-110
36. Daker K A, Lehrer R I, Ganz T. Isolation and characterization of human defensin cDNA clone. Pro Natl Acad Sci USA. 1989, 86(1): 342
37. Del P, Bonitto M, Zuccon D, Cervella P, Spano A, Amoroso A, Crovella S. β-defensin 1 gene variability among non-human primates. Immunogenetics. 2002, 53: 907-913
38. Diamond D L, Kimball JR, Krisanaprakomkit S, et al. Detection of β-defensins secreted by human oral epithelial cells. Journal of immunological methods. 2001, 256: 65-76
39. Diamond G, Zasloff M, Eck H, et al. Tracheal antimicrobial peptide, a novel Cysteine-rich peptide from mammalian tracheal nucosa: peptide isolation and cloning of cDNA. Pro Natl Acad Sci USA. 1991, 88(10): 3952-3956
40.Ding M(丁鸣),Hellekant G,Hu Zhong(胡忠).Characterization and chemicalmodifization of brazzein, a high potency thermostable sweet protein from pentadiplander brazzeana(耐热甜味蛋白Brazzein的特性和化学修饰).acta Botanica-Yunnanica(云南植物研究).1996,18(2):123～133
41. Donovan RS, Robinson CW, Glick BR. Review: optimizing inducer and cultrue condictions for expression of foreign proteins under the control of the Lac promoter. Journal of Industrial Microbiology. 1996, 16: 145-154
42. Duits L A, Rademaker M, Ravensbergen B, et al. Inhibition of hBD3, but not hBD-1 and hBD-2, mRNA expression by corticosteroids. Biochem Biophys Res Commun. 2001, 280(2): 522—555
43. Edens L, Bom I, Adrianus M, Ledeboer. synthesis and processing of the plant protein Thaumatin in Yeast. Cell. 1984, 37: 629-633
44. Edens L, Heslinga L, Klok R, et al. cloning of cDNA encoding the sweet tasting plant protein thaumatin and its expression in Escherichia coil Gone. 1982, 18: 1-2
45. Epand R M, Vogel H J. Diversity of antimicrobial peptides and their mechanisms of action. Biochim Biophys Acta. 1999, 1462: 11-28
46.Fan Changsheng(范长胜),甜蛋白的开发与应用研究.食品与发酵工业.2001,27(12):50-54
47. Fang X, Peng L, Xu Z, Wu J, Cen P. Cloning and expression of human beta-defensin-2 gene in Escherichia coil Protein Peptide Letter. 2002, 9(1): 31-7
48. Fariba M, Assadi-Porter, David J, Aceti, et al. sweetness determinant site of Brazzein, a small, heat-stable, sweet-tasting protein. Archieves of Biochemistry and Biophysics. 2000, 376(2): 259-265
49. Fariba M, Assadi-Porter, David J, Aceti, et al. efficient production of recombinant Brazzein, a small, heat-stable, sweet-tasting protein of plant orgin. Archieves of Biochemistry and Biophysics. 2000, 376(2): 252-258
50. Fans I. recent developments in the characterization and biotechnological production of sweeting-tasting proteins. Appllied Microbiological Biotechnology. 2000, 53: 145-151
51. Faus I, Patino Cristina, Del RJL, et al. expression of a synthetic gene encoding the sweet-tasting protein thaumatin in Escherichia coil Biochem Biophysi Resear Commu. 1996, 229: 121-127
52. Faus I, Del Moral C, Adroer N. secretion of the swotting-tasting protein thaumatin by recombinant strains of Aspergillus niger va Awamori. Appl. Microbiol Biotechnol. 1998, 49: 393-398
53. Faus I, Patino C, Del Rio J L, et al. expression of a synthetic encoding the sweet-tasting protein thaumatin in the filamentous fungus Penicillium roquewfortii. Biotechnology Letter. 1997, 19: 1185-1191
54. Frohlicho Poc, Young L G. Organization of human gene encoding the epididymis-specific EP2 protein variants and its relationship to defensin genes. Biol Report. 2001, 64: 1072-1079
55. Frye M, Bargon J, Gropp R. Expression of human beta defensin-1 promotes differentiation of keratinoeytes. J. Mol. Med. 2001, 79(5-6): 275-282
56. Ganz T, Selsted M E, Szklarek D, et al. Defensins, Natural peptide antibiotics of human neutrophils. The Journal of Clinical Investigation. 1985 76(4): 1427-35
57. Ganz T, Lehrer R I. Defensins, Current Opinion in Immunology. 1994, 6: 584-589
58. Gao Guang-Hua, Dia J X, Ming D, et al. solution conformation of brazzein by ~1H nuclear magnetic resonance: resonance assignment and secondary structure. International Journal of Biological Macromolecules. 1999, 24: 351-359
59.Gao SH N(高世年)主编.《实用食品添加剂》.天津科学技术出版社.2000,197-207
60. Garcia J R, Jaumann F, Schultz S, Krause A, et al. Identification of a noval, multifunctional β defensin (human β defensin 3 )with specific antimierobial activity. Cell tissue Res. 2001, 306: 257-264
61. Garcia J R, Krause A, Schulz S, et al. Human β-defensin-4: a novel inducible peptide with a special salt-sensitive spectrum of antimicrobial activity. The FASEB Journl. 2001, 15: 1819-1821
62. Garcia JR, Jaumann F, Schulz S, et al. Identification of a noval, multifunctional beta-defensin(human beta-defensin 3) with special antimicrobial activity, its interaction with plasma membrance of Xenopus oocytes and the induction of macrophgge chemoattraction. Cell Tissue Res. 2001, 306(2): 257-264
63. Goochee C F, Gramer M J, Andersen D C, Bahr J B, Rasmussen J R. The oligosaccharides of glycoproteins: bioprocess factors affecting oligosaccharide structure and their effect on glycoprotein proteins. Biotechnology. 1991, 9: 1347-1355
64.Guo L D(郭良栋),Qian Sh J(钱世钧),Ye J(叶军),Jiao Q H(矫庆华).Studies on the culture conditions of engineered strain of human lysozyme(人溶菌酶工程菌培养条件的研究).Acta Microbiologica Sinica(微生物学报).1997,37(1):53-57
65.Guo M J(郭美锦),Zhumag Y P(庄英萍),et al.Expression of recomibinant human serum albumin in genetically enginnred pichia pastoris in high-density fermentation(重组巴氏毕赤酵母高密度发酵表达rHSA).Acta Microbiologica Sinica(微生物学报).2002,42(1):62-68
66. Hancock REW, Falla T and Brown MH. Cationic bacteriecidal peptides. Adv Microb Physiol. 1995, 37: 135-175.
67. Hancock Robert EW. Cationic peptides: effectors in innate imminity and novel antimicrobials, the Lancet Infectious Dieases. 2001, 11(1): 156-164
68. Hancock REW, Chapple D S. peptide antibiotics. Antimicrob Agents Chemother. 1999, 43: 1317-1323
69. Harder J, Bartels J, Christophers E, Schroder JM. A peptide antibiotic from human skin. Nature. 1997, 387 :861
70. Harder J, Bartels J, Christophers E, Schroder JM. Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic. Journal of Biological Chemisity. 2001, 276(8): 5707-5713
71. Hasslacher M, et al. Hight-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree hevea brasiliensis in microbial hosts. Pro Expr Purif. 1997, 11, 61-71
72. Haynes R J, Tighe P J, Dua H S. Antimicrobial defensin peptides of the human ocular surface. Br f Ophthalmol. 1999, 83: 737-741
73.He Cheng(何诚),Zhu Yunsong(朱运松).甲醇营养型酵母表达系统的研究进展.生物工程进展.1998,18(3),7-11
74. Hoffmann JA, Hertu C. Insect defensins: inducible antibacterial peptides. Immunol Today. 1992, 13(10): 411-415
75. Hoover D, Wu Zhibin, Tucker K, Lu W, Lubkowski J. Antimicrobial characterization of human β-defensin 3 derivatives. Antimicrobial agent and chemotherapy. 2003, 9: 2804-2809
76. Hoover D, Rajashankar K R, et al. the structure of human β-defensin-2 shows evidence of higher order oligimerization. The journal of biological chemistry. 2000, 275(42): 32911-32918
77.Hu Zhong(胡忠),He ming(何敏).study on Mabinlin, a sweet protein from the seeds of capparis masaikai levi Ⅰ extraction, purification and certain characteristics(吗槟榔甜蛋白的研究Ⅰ提取、纯化和某些特性).Acta Botanica yunnanica(云南植物研究).1983,5(2):207-212
78.Huang Yadong(黄亚东),Zheng Qi(郑青),et al.Modification of Cecropin AD gene and its expression in Pichia pastoris. Journal of China university of technology(Natural science edition)(华南理工大学学报,自然科学版).2002,30(2):13—16
79.Huang Yin(黄音),Liu Feipeng(刘飞鹏),Zhou Tianhong(周天鸿),Zhu Jiaming(朱嘉明).蛙皮素—蜂毒素杂合肽基因在大肠杆菌的克隆与诱导表达.生物工程学报.2001,17(2):207-210
80. Illingworth C, Larson G Hellenkant G secretion of the sweet-tasting protein Thaumatin by Bacillus Subtilis. Biotechnology Letter. 1988, 16: 587-592
81. Ishkawa K, Ota M, Ariyoshi Y, Sasaki H, et al. Crystallization and preliminary X-ray analysis of brazzein, a new sweet protein. Acta Crystallogr D. 1996, 52: 577-578
82. Izawa H, Ota M, Kohmura M, Ariyoshi Y. Synthesis and characterization of the sweet protein brazzein. Biopolymers. 1996, 39(1): 95-101
83. Jia H P, Schutte B C, Schudy A, et al. Discovery of new human beta-defensins using a genomics-based approad. Gene. 2001, 263: 211-218
84.Jiang Y(江阳),Shi H Zh(时宏珍),et al.Study on the expression of recombination of human Interleukin-13(rhIL-13) in methylotropic yeast pichia pastoris and its effect on the induction of DC to differentiate in vitro.中国免疫学杂志.2003,19:82—87
85. Jin Z, Danilova V, Assadi-Porter FM, Aceti DJ, Markley JL, Hellekant G Critical regions for the sweetness of brazzein. FEBS Lett. 2003, 544(1-3): 33-7
86. Jin Z, Danilova V, Assadi-Porter FM, Markley JL, Hellekant G. Monkey electrophysiological and human psychophysical responses to mutants of the sweet protein brazzein: delineating brazzein sweetness. Chem Senses. 2003, 28(6): 491-8
87. Joly S, Maze C, Mccray P B, Guthmiller J M. Human β-defensin 2 and 3 demonstrate strain-selective activity against oral microorganisms. J Clin Microbiol. 2004, 3: 1024-1029
88. Jones DE, Bevins CL. Paneth cell of the human small intestine express an antimicrobial peptide gene. J boil Chem. 1992, 267: 23216-23215
89. Jones DE, Bevins CL. Defensin-6 mRNA in human Paneth cell: implications for antimicrobial peptides in hose defensin of the human bowel. FEBS Letter. 1993, 315: 187-192
90. Kapralek F, Jecmen P, Sedlacek J, Fabry M, Zadrazil S. Fermentation conditions for high level expression of tac-promoter-controlled calf prochymosin cDNA in E.coli HB101. Biotechnology and bioengibeering. 1991, 37: 71-79
91. Keneth JM, Roert IL. β-sheet antibiotic peptic as potential detal therapeutics. International Journal of Antimicrobial Agents. 1998, 9: 269-280
92. Kim SH, Kang CH, et al. redesingniting a sweet protein: increased stability and renaturability. Protein Engineering. 1989, 2(8): 571-579
93. Kohmura M, Nio N, Ariyoshi Y. Highly probable active site of the sweet protein monellin. Biosci Biotechnol Biochem. 1992, 56(12): 1937-42
94. Kondo K, Miura Y, Sone H, et al. high-level expression of a sweet protein monoeilin, in the food yeast Candida utilis. Nat Biotechnol. 1997, 15: 453-457
95.Kong J Q(孔建强),Zhao Q(赵琦),Gao Y(高音),et al.Sweet protein thaumatin and it's genetic engineering(奇异果甜蛋白及其基因工程).Hereditas(beijing)(遗传).2003,25(2):232—236
96. Koutz P, Davis GR, Stillman C, Barringer K, et al. Structural comparison of the Pichia Pastoris alcohol oxidase genes. Yeast. 1989, 5: 167-177
97. Krisanaprakornkit S, Weinberg A, Perez CN, et al. Expression of the peptide antibiotic human beta-defensin 1 in cultured gingival epithelial cell and gingival tissue. Infection and immunity. 1998, 66: 4222-4228
98. Lee SY, Lee JH, Chang HJ, Cho JM, Jung JW, Lee W. Solution structure of a sweet protein single-chain monellin determined by nuclear magnetic resonance and dynamical simulated annealing calculations. Biochemistry. 1999, 38(8): 2340-6.
99. Lehrer RI, Barton A, Dither KA. Interaction of human defensins with E.coli mechanism of bactericidal activity. J Ciin Invest. 1989, 84(2): 553-561
100. Lehrer RI. defensin: antimicrobial and cytotoxic peptides of mammalian cells. A.Rev Immun. 1993, 11: 105-128
101. Lehrer RI, Ganz T. Antimicrobial peptides in mammalian and insect host defence. Cur opin Immu. 1999, 11: 23-27
102. Lehrer RI, Ganz T. Endogenous vertebrate antibiotics defensins, protegrins and other cysteine-rich antimicrobial peptides. Ann NY Acad Sci. 1996, 797: 228-239
103.Li Guangkuo(李广阔),Li X(李旭),Tian F Zh(田伏洲),et al.人源性空回肠粘膜防御素-5的基因的克隆.中华实验外科杂志.2003,20(10):884-885
104.Li Hongiun(李洪军).防御素研究进展.生物工程进展.2001,21(3):34-38
105.Li Hongzhao(李洪钊),Li Liangzhu(李亮助),Sun Qiangming(孙强明),et al.Strategies for optimizing pichia pastoris expression syetem(巴斯德毕氏酵母表达系统优化策略).Acta Microbiologica sinica(微生物学报).2003,43(2):288-292
106.Li G L(李广林),Guo A G(郭蔼光),Fan S H(范三红).Cloning and expression of brazzein gene(植物Brazzein基因的克隆与表达).Acta Bol Boreal-Occident Sin(西北植物学报).2004,24(7):1271-1274
107.Li Jing(李晶),Zhao Xiaoxiang(赵晓样).甲醇酵母基因表达系统的研究进展.生物工程进展.1999,19(2),17-20
108.Li X D(李晓东),Liu J M(刘敬梅),Chen H(陈杭),et al.Genetic transformation of sweet protein MBLⅡ on tomato(甜蛋白基因MBLⅡ对番茄的遗传转化).Acta Bol Boreal-Occident Sin(西北植物学报).2004,24(5):808-811
109.Li Y(李英),Yin J(尹俊),Zhang Sh J(张树军).Synthesis and cloning of cDNA encoding the sweet plant protein monellin(植物甜蛋白monellin基因的合成和克隆).Journal of Inner Mongolia Agricultural University(内蒙古农业大学学报).2001,22(3):35-38
110. Liao YF, Lal A, Moremen KW. Cloning, expression, purification and characterization of the human broad specificity lysosomal acid alpha-mammosodase. Journal of biology and chemistry. 1996, 271, 28348-28358
111. Lichtenstein A. Mechanism of mammalian cell lysis mediated by peptide defensins, Evidence for an initial alternation of the plasma membrance. J Clin Invest. 1999, 88(1): 93-100
112.Ling GD(凌关庭)主编.《食品添加剂手册》(第三版).化学工业出版社.2003,645-666
113. Linzmeier R, Miehaelson D, Liu L, et al. The structure of neutrophil defensin genes. FEBS Letter. 1993, 321: 267-273
114. Linzmeier R, Ho C H, Hoang BV, Ganz T. a 450-kb of defensin genes on human chromosome 8P23. gene. 1999, 233: 205-211
115.Liu J M(刘敬梅),Chen D M(陈大明),Chen H(陈杭).Genetic transformation and plant regeneration of lettuce with sweet protein gene MBLⅡ(甜蛋白基因MBLⅡ对莴苣的遗传转化).Acta Horticulturae Sinica(园艺学报).2001,28(3):246-250
116. Liu L, Wang L, Jia H P, et al. Structure of the mapping of the human β-defensin HBD-2 gene and its expression at sites of inflammation. Gene. 1998, 222: 237-244
117.Liu Q Y(刘秋云),He ZH M(贺竹梅),Cao J(曹俊),et al.利用农杆菌系统将超甜蛋白基因导入烟草.遗传.1998,20(1):24-27
118. Liu W C, Zhang XY, Hu JL, tumor cell eytolysis by defensins of human neutrphils in vitro. Inter J Modern cancer ther. 1998, 1: 45
119.Liu W C(刘文超),Mu H X(穆怀兴),Ren J(任军),Zhang X Y(张学庸),et al.防御素对胃癌细胞系体外的杀伤作用.World Chin J Digestol(世界华人消化杂志).2001,9(6):622-626
120. Liu X, Hu Z, Maeda S, Aiuchi T, Nakaya K. Purification, complete amina acid sequence and structure chaeterization of the heat stable sweet protein, mabinlin Ⅱ. Eur J Bioehem. 1993, 211: 281-287
121.Lu B(陆兵),Xia J(夏杰),Xu D Sh(徐殿胜),Chen G H(陈国豪).The Lactose induction for the expression of rhNTA under the control of φ10 promoter in E. coli(φ10启动子控制下的大肠杆菌中rhNTA表达的乳糖诱导).Journal of East China university of science and technology(华东理工大学学报).2001,4:131-134
122.Ma li(马骊),Wang Xiaoning(王小宁),Zhang Zhiqing(张智清),et al.Expression, Purification and biological activity analysis of human vascular endothelial growth factor (VEGF_(165)) in pichia pastoris(人血管内皮因子(VEGF_(165))在毕氏酵母中的表达及生物活 性分析).Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(3):325-330
123. Mackewicz C E, Yuan J, Tran P, et al. alpha-defensins can have anti-HIV activity but are not CD8 cell anti-HIV factors. AIDS. 2003, 17(14): 31
124. Malsetta G, Batoni G, Esin S, et al. Activity of Human β-defensins 3 alone or comibined with other antimicrobial agent against oral bacteria. Antimicrobial agent and chemotherapy. 2003, 10, 3349-3351
125. Makrides SC, strategies for achieving high-level expression of genes in E.coli. Microbiological review. 1996, 60(3): 512-516
126. Mallow EB, Harris A, Salzman N, et al. Human enteric defensins. gene struction and developmental expression. J Biol Chem. 1996, 271 (8): 4038-4045
127. Masuda T, Tamaki S, Kaneko R, Wada R, Fujita Y, Mehta A, Kitabatake N. Cloning, expression and characterization of recombinant sweet-protein thaumatin Ⅱ using the methylotrophic yeast Pichia pastoris. Biotechnoi Bioeng. 2004, 85(7): 761-9.
128. Masuda J A, Nirasawa S, Nakaya K, Kurihara Y. Cloning and sequencing of a cDNA encoding a taste-modifying protein miraculin. Gene. 1995, 161: 175-177
129. Mathews M, Jia H P, Guthmiller J M, et al. Production of beta-defensin antimicrobial peptides by oral mucosa and salivary gland. Infection and Immunity. 1999, 67: 2740-2745
130. Mineshiba F, Takashiba S, Mineshiba J, Matsuura K, Kokeguehi S, et al. Antibacterial activity of synthetic human β-denfensin 2 against periodontal bacteria. J Int Acad Periodontal. 2003, 5: 35-40
131. Ming D, Hellekant G, Brazzein. a new high potency sweet protein from Pentadiplandra brazzeana Baillon. FEBS Letter. 1994, 355: 106-108
132. Moralejo FJ, Cardoza RE, Gutierrz S, et al. Thaumtin production in Aspergillus awamori by use of expression cassettes with strong fungal promoters and high gene dosage. Applied and Environmental Microbiology. 1999, 65 (3): 1168-1174
133. Moralejo F J, Cardoza R E, Gutierrz S, et al. overexpression and lack of degradation of thaumatin in an aspergillopepsin A-defective mutant of Aspergillus awamori containing an insertion in the pepA gene. Appl Microbiol biotechnol. 2000, 54: 772-777
134. Morris J A, Cagan R H. purification of Monellin, the sweet principle of Discoreophyllum curarainisii. Biochem. Biophys Acta. 1972, 261: 114-122
135. Muller C, Markovie-Lipkovski J, Klatt T. human α-defensins HNPs-1,-2,and-3 in renal cell carcinoma influences on tumor cell proliferation. American journal of patbology. 2002, 160(4): 1311-1323
136. Nirasawa S, Nishino T, Katahira M, et al. Struction of heat-stable and unstable homologues of the sweet protein mabinlin. Eur J Biochem. 1994, 223: 989-995
137. Ogawa M, Nakamura S, Atsuchi T, et al. Macromolecu lar antimicrobial glycoprotein, achacin, expressed in a methylotrophic yeast Pichia Pastoris. FEBS Letters. 1999, 448: 41-44
138. Ohtani W, Nawa Y, Takeshima K, Kamuro H, kobayashi K, Ohmura T. Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia Pastori. Anal Biochem. 1998, 256, 56-62
139. Ortega M R, Ganz T, Milner S M. Human beta defensin is absent in bum blister fluid. Burns. 2000, 26: 724-726
140. Periathamby AR, Andrew RD. Current status of defensins and their role in innate and adaptive immunity. FEMS Microbiology Letters. 2002, 206: 9-18
141. Pang SK, Oberhans SM, Rasmussen J K, et al. Expression of gene encoding a scorpion insectotoxin peptide in yeast, bacteria and plants. Gene. 1992, 116: 165-172
142. Penarrubia L, Kim R, Giovannoni J. production of the sweet protein monellin in transgenic plants. Biotechnol. 1992, 10: 561-564
143. Peng L, Xu Z, Fang X, Wang F, Yang S, Cen P. Preferential codons enhancing the expression level of human beta-defensin-2 in recombinant Escherichia Coli. Protein Pept Lett. 2004, 11(4): 339-344
144. Peng Yu-Cai, et al. Production of active polyomavirus large T antigen inyeast Pichia Pastoris. Virus Reasearch. 1997, 49: 41-47
145.Peng Li(彭力),Xu Zhinan(徐志南),et al.Advances in the research of human defensins. Chinese Journal of biotechnology(生物工程学报).2003,19(3):261-266
146.Peng Yi(彭毅),Bu Wei(步威).甲醇酵母表达系统.Biotechnology Information(生物技术通报).2000,1,38-41
147.Peng Yi(彭毅),Yang Xicai(杨希才),Kang liangyi(康良仪).影响甲醇酵母中外源蛋白表达的因素.Biotechnology Information(生物技术通报).2000,4:33-36
148. Peschel A, Ralph W. Jack, Michael Otto. Staphylococcus aureus Resistance to Human Defensins and Evasion of Neutrophil Killing via the Novel Virulence Factor MprF Is Based on Modification of Membrane Lipids with L-Lysine. The Journal of Experimental Medicine. 2001, 193(9): 1067-1076
149. Piers K L, Brown M H, Hancock REW. Recombinant DNA procedures for producing small antimicrobial cationic peptide in bacteria. Gene. 1993, 134(1): 7-13
150. Prohaszka Z, Nemet K, Csermly P, et al. defensins purified from human granuloeytes bind C1q and activate the classical complement pathway like the transmembrane glyeoprotein gp41 of HIV-1. Molecular immunology. 1997, 34(11): 809-816
151.Qu X D(曲晓丹),Wang Ai X(王爱霞).人中性粒细胞防御素的提纯及活性分析.中华医学杂志.1991,71(11):616—619
152. Quinones-mateu M, Lederman M M, Feng Z, et al. Human epithelial-defensin 2 and 3 inhibit HIV-1 replication. AIDS. 2003, 17(16): 39-43
153. Raj PA, Antonyraj KJ, Karunakaran T. Large-scale synthsis and functional elements for the antimicrobial activity of defensins. Biochem J. 2000, 347: 633-64
154. Reiehart J M, Petit I, legtain M, et al. expression an inducible antibacterial peptide from the fleshly Phormia terranovae. Invert reproduct develop. 1992, 21 (1): 15-24
155.Ren H Y(任洪艳),Zhang L H(张苓花),Wang Y J(王运吉).Cloning of human α-defensin 1 and iu structural analysis(人α-防御素HNP-1基因克隆及其结构分析).Journal of Dalian institute of light industry(大连轻工学院学报).2003,22(4):265-269
156. Risso A. Leukocyte antimicrobial peptide: multifunetional effector molecules of innate immunity. J Leukocyte Biol. 2000, 68: 785-792
157. Robert IL, Tomas Ganz. Defensins of vertbrate animal. Current Opinion in Immunology. 2002, 14: 96-102
158. Romanos MA, Clare JJ, Beesley KM, Rayment FB. Recombinant Bordetella Pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties. Vaccine. 1991, 9: 901-906
159. Romanos Mike, Wellcome Glaxo, Beckenham. Advances in the use of pichia pastoris of high level gene expression. Current Opinion in Biotechnology. 1995, 6: 527-533
160. Sahly H, Schubert S, Harder J, et al. Bukholderia is highly resistant to human beta-defensin 3. Antimierobial agents and chemotherapy. 2003, 5: 1739-1741
161. Sambrook Joseph, Russell David W. Molecular Cloning: A laboratory Manual. Cold Spring Harbor Laboratory Press. 2001
162. Sehroder JM. Epithelial peptide antibiotic. Biochem Pharma. 1999, 57: 121-134.
163. Sehroder JM, Harder J. Molecules in focus human beta-defensin-2. The international journal of biochemistry & cell biology. 1999, 31: 635-651
164. Scorer CA, Clare JJ, McCombin WR, et al. Rapid selection using G418 of high copy number transformants of Pichia Pastoris for high-level foreign gene expression. Bioteehnoiogy. 1994, 12: 181-184
165. Selsted ME. Primary struction of three human neutrophlis defensins. The Journal of Clinical Investigation. 1985, 76: 1436-40
166. Selsted M E, Tang Y Q, Morris W L, Mcguire P A, et al. Purification primary structures and antibacterial activities of β-defensins, a new family of antimicrobial peptides from bovine neutrphils. J Biol Chem. 1993, 268: 6641-6648
167. Seo SJ, Ahn SW, Hong CK, Ro Bl. Expression of β-defensins in human kerationoeyte cell lines, Journal of dermatological science. 2001, 27: 183-191
168.Shao L L(邵玲莉),Zhou N D(周楠迪),Jin J(金坚),et al.Optimization of culture medium for the production of recombinant human angiostatin(重组人血管抑制素生产菌培养基优化研究).Pharmaceutical biotechnology(药物生物技术).2004,11(2):91—95
169.Shao L L(邵玲莉),Zhou N D(周楠迪),Wu Q F(吴奇凡),Jin J(金坚),et al.Fermentation contion of angiostatin by recombinant(重组大肠杆菌生产人血管抑制素的发酵条件).journal of Wuxi university of light industry(无锡轻工大学学报).2004,23(2):8—11
170. Sharma S, Verma I, Khuller GK. Therapeutic potential of human neutrophil peptide 1 against experimental tuerculosis. Antimicrobial agent and chemotherapy. 2001, 2: 639-640
171. Sharp PM, Tuohy TMF, Mosurski KR. Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes. Nuleid acid Research. 1986, 14(13): 5215-5143
172. Shen S, Sulter G, Jeffreies T W, et al. A strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast pichia pastoris. Gene. 1998, 216: 93-106
173.Shiqiang(石强),Liu Feipeng(刘飞鹏).抗菌肽基因的表达和转基因研究现状.生物工程进展.2000,20(11):37—40
174. Slootstra JW, De Geus P, Haas H, Verrips CT, Meloen RH. Possible active site of the sweet-tasting protein thaumatin. Chem Senses. 1995, 20(5): 535-43
175. Somoza JR, Cho JM, Kim SH. The taste-active regions of monellin, a potently sweet protein. Chem Senses. 1995, 20(1): 61-8.
176. Sreekrishna K, Brankamp RG, Clare JJ, et al. strategies for optimal synthesis and secretion of heterogous proteins in the methylotrophic yeast pichia pastoris. Gene. 1997, 190: 55-62
177. Spadaccini R, Crescenzi O, Tancredi T, De Casamassimi N, et al. Solution structure of a sweet protein: NMR study of MNEI, a single chain monellin. J Mol Biol. 2001, 305(3): 505-14
178. Suami T, Hough L, Machinami T, et al. Molecular mechanisms of sweet taste: The sweet protcin, Thaumatin I. Food chemistry. 1997, 60(3): 277-285
179.Tang Bing(唐彬),Huang Ning(黄宁),Wu Qi(吴琦),et al.Studies on the expression of transferred human α defensin HNP1 gene in tracheal eoithelial cells cultivated in vitro. Chinese Journal of Microbiology(中华微生物学和免疫学杂志).1999,19(6):481-484
180.Tuo xiaoye(庹晓晔),Xu Mingda(徐明达),Chen Bi(陈璧),et al.Fusion expression of human beta-defensin 3 in E.coli and primary analysis of its antimicrobial activities. Bull Acad Mil Sci(军事医学科学院院刊).2004,28(2):114-122
181. Urabatsch IL, Wilke MS, Gimi K, et al. Purification and characterization of N-glycosylation mutant mouse and human P-glycoproteins expressed in pichia pastoris cells. Arch Biochem Biophys. 2001, 388: 171-177
182. Valore EV, Park C H, Quayle A J, et al. Human beta-defensin-1: an antimicrobiai peptide of urgenital tissues. Journal of clinical investigation. 1998, 101 (8): 1633-1642
183. Van del wel H. Structure modification of sweet proteins and its influence on sensory properties. Chemistry and Industry. 1983, 3: 19
184. Van del wel H, lyengar RB. Assignment of the disulfide bonds in the sweet tasting protein thaumatin I. Eur J Biochem. 1984, 144: 41
185. Van der Wel H, Larson G. Isolation and charcterization ofpentadin, the sweet principle of Pentadiplandra brazzeana Baillon. Chem Senses. 1989, 14: 73-79
186. Van der Wel H, Loeve K. Isolation and charcterization of Thaumatin Ⅰ and Ⅱ, the sweet tasting protein from Thaumatotoccus daniellii Benth. Eur J Biochem. 1972, 31: 221-225
187. Valore EV, Park CH, Quayle AJ, et al. Human beta-defensin 1: an antimierobial peptide of urogenital tissue. Journal of clinical investigation. 1998, 101(8): 1633-1642
188. Wada A, Ogushi K, Kimura T, et al. Helicobaeter pylori-mediated transcriptional regulation of the human beta-defensin 2 gene requires NF-kappaB. Cell Mierobiol. 2001, 3 (2): 115-123
189. Wade D, Boman A, Waklin. amino acid-containing channel-forming antibiotic peptides. Proc.Natl Acad Sci USA. 1990, 87: 4761-4765
190. Wang F, Fang X, Xu Z, Peng L, Cen P. Fusion expression of human beta-defensin-2 from multiple joined genes in Escheriehia coli. Prep biochem Bioteehnol. 2004, 34 (3): 215-225
191.Wang Tongying(王同映),Wang Changmei(王昌梅),Wei Guang(魏光),et al.Expression of the recombinant human Intedekin-Ⅱ in pichia pastoris(重组人白细胞介素Ⅱ在毕氏酵母中的表达).Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(6):659-664
192.Wang Weirong(王维荣),Liu Aiming(刘爱民),Qian Zhikang(钱志康),Huang Weida (黄伟达).青岛海葵强心活性多肽在毕氏酵母中的分泌表达.生物化学与生物物理进展.2000.27(4):407-411
193.Wang Yanru(王艳茹),Ren Hongwei(任宏伟),et al.人三叶因子3在毕赤酵母中的表达及活性分析.生物化学与生物物理学报.2001,33(6):653-658
194. Waterham HR, Digan ME, et al. Isolation of the pichia pastoris glyceraldehydes-3-phosphate dehydrogenase gene and regulation and use of its promoter. Gene. 1997, 186: 37-44
195. Weichert MJ, Pagratis M, Curry SR, Blackmore R. stabilization of apoglobin by low temperature increases yield of soluble recombinant hemoglobin in E.coli. Applied and environmental microbiology. 1997, 11: 4313-4320
196. Wimley WC, Seleted ME, White SH. Interactions between human defensins and lipid bilayers: evidence for formation of multimeric pores. Protein Science. 1994, 3: 1362-1373
197. Withers-Martinez C, carpenter EP, Hackett F, et al. PCR-based gene synthesis as an efficient approach for expression of the A+T-rich malaria genome, protein engineer. 1999, 12: 113-1120
198. Witty M. propeothaumatin Ⅱ is processed to biological activity in solanum tuberosum. Biotechnol Letter. 1990, 12: 131-136
199.Wu G(吴刚),Wang Y M(汪亚平),Zhu Z Y(朱作言).Secretion expression of Monellin in Pichia Pastoris(甜蛋白Monellin在毕赤酵母中的分泌表达).高技术通讯.2003,11:20-23
200. Wu M, Maier E, Bena R, Hancock REW. Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemstry. 1999, 38: 7235-7234
201.Xin Li(辛利),Zhang Li(张励),et al.人血管抑制素在毕赤酵母中的表达及活性测定.Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(3):291-295
202.Xiong Aisheng(熊爱生),Peng Rihe(彭日荷),Li Xian(李贤).Influence of signal peptide sequences on the expression of heterogeneous proteins in pichia pastoris(信号肽序列对毕氏酵母表达外源蛋白质的影响).Acta Biochimica et Biophysica Siniea(生物化学与生物物理学报).2003,35(2):154-160
203. Xiong L, Sun S. Molecular cloning and transgenic expression of sweet protein mabinlin in potato tubers. Plant Physiol. 1996, 111: 57
204.Xu ZH N(徐志南),Liu G(刘刚),Cen P L(岑沛霖).Research on fermentation for hEGF production in recombinant Escherichia coli(重组大肠杆菌生产hEGF发酵条件的研究).Journal of Zhejiang universiq4engineering science)(浙江大学学报(工学版)).2001,35(1):25-28
205.Yang Ch L(杨成丽),Liu Sh N(刘树楠),Zhou J Y(周吉源),et al.Introduction of Thaumatin gene into Tobacco(Nicotiana tabacum L)through Agrobacterium-mediated genetic transformation(高效烟草遗传转化体系的建立及其甜蛋白基因的导入).Biotechnology(生物技术).2004,14(2):9-11
206. Yang D, Biragyn A, Kwak L W. Oppenheim J J, Mammalian defensins in immunity: more than just microbicibal. Trends Immunol. 2002, 23: 291-296
207. Yang D, Chertov D, et al. beta-defensin: linking innate and adaptive immunity through dentritic and T cell CCR6. Science. 1999, 286: 525-528
208. Yang D, Chen Q, Chertov O, et al. Human neutrophil defensins selectively chemoattract naive T and immature dendritic cells. J leukoc Biol. 2000, 68: 9-14
209.Yang M ZH(杨美珠),Pan N S(潘乃楗),Chen ZH L(陈章良).高效马铃薯遗传转化体系的建立及甜蛋白基因的导入.植物学报.1992,34(1):31-36
210.Yang X J(杨晓娟),Chen J(陈静),Ma X R(马欣荣),et al.人β-防御素-2植物表达载体的构建.四川大学学报(自然科学版).2002,39:55-59
211.Yao Bing(姚斌),Zhang Chunyi(张春义),Wang jianghua(王建华),Fan Yunliu(范云六).高效表达具有生物活性的植酸梅的毕氏酵母.中国科学(C辑).1998,28(3):237—243
212.Ye J(叶娇),Chen Ch H(陈长华),Yang Y Q(杨雅琴),et al.expression of recombinant human superoxide dismutase gene in E.coli induced by Lactosc(乳糖诱导重组人SOD基因 在大肠杆菌中的表达).Chinese journal of pharmaceuticals(中国医药工业杂志).2001,32(2):536-539
213.Yu Xuehong(于学红),Yuan Hanying(袁汉英),et al.Expression of Monellin, a sweet protein, in pichia pastoris. Journal of Fudan University(Natuml science)(复旦学报 自然科学版).2002,41(6):650-654
214.Yuan Hanying(袁汉英),Ai HS(艾华水),Fan ZS(范长胜)et al.Construction of the expression vector for single-chain Monellin gene and its expression(单链莫热宁基因表达载体的构建及其分泌表达).Journal of Fudan University(Natural science)(复旦学报 自然科学版).2003,42(4):
215. Zhang L, Falla T, Wu M, et al. Determination of recombinant production of antimicrobial cationic peptides and creation of peptide variants in bacteria. Biochemical and Biophysical Research Commutation. 1998, 247: 674-680
216.Zhang Chunyi(张春义),Zhan jun(赵军),Wu Changjian(武长剑),Fan Yunliu(范云六).利用毕氏酵母表达抗真菌鳖素基因.科学通报.1998,43(19):2085-2089
217. Zhang G L, Wu H, Shi J. Molecular cloning and tissue expression of procine β-defensin-1. FEBS Letter. 1998, 424: 37-40
218.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).Expression of human defensin —1(HNP-1)in E.coli. Chinese biochemical journal(生物化学杂志).1997,13(3);264-269
219.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).human defensin 1(HNP-1) expression in Tobacco: A preliminary observation on anti-TMV activity of transgenic Tobacco plants. Acta Botaniea sinica(植物学报).1997,39(6):574-576
220.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).Transient expression on human defensin 1(HNP-1)in Tobacco protoplast. Acla Biologiae experimentalis sinica(实验生物学报).1997,30(4):209-211
221.Zhang Yi(张毅),Qu X M(屈贤铭),Yang Sh L(杨胜利).High cell density culture of the recombinant E.coli harboring h GM-CSF/IL-3 fusion expression plasmid(重组GM-CSF/IL-3融合蛋白表达菌发酵条件的研究).Industrial Microbiology(工业微生物).2000,30(2):8-11
222.Zhao Donghong(赵东红),Dai Zhuying(戴祝英),Zhuo Kaiya(周开亚).昆虫抗菌肽的功能、作用机理与分子生物学研究最新进展.生物工程进展.1999,19(5):14—18
223. Zhao C Q, Wang I, Lehrer R I. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Letters. 1996, 396: 319-322
224.Zhou J H(周家华),Cui Y D(崔英德),Li B N(黎碧娜)等主编.《食品添加剂》.化学工业出版社.2001
225.Zhou P(周平),Pan N S(潘乃稽),et al.外源甜蛋白Thaumatin基因转入烟草的研究.植物学通报.1994,11(4):17-20
1. Andersson SGE, Kurland CG. Coden preferences in free-living microorganisms. Microbiological review. 1990(6), 198-210
2. Assadi-porte FM, Aceti DJ, Cheng H, Markley J L. Efficient production of recombinant Brazzein, a small, heat-stable, sweet-tasting protein of plant origin. Archives of Biochemistry Biophysics. 2000, 376(2): 252-258
3. Assadi-porter F M, Aceti D J, Markley J L. Sweetness determinant sites of Brazzein, a small, heat-stable, sweet-tasting protein. Archives of Biochemistry Biophysics. 2000, 376(2): 259-265
4. Antcheva N, Bonitto M, Zelezetsky I, et al. Effects of positively selected sequence variations in human and Macaca fascicularis β defensin 2 on antimicrobial activity. Antimicrob Agents Chemother. 2004, 2: 685-688
5. Bals R, Wang X, Wu Z, et al. Human β-defensin-2 is a salt-sensitive peptide antibiotic expressed in human lung. Journal of clinical investigation. 1998, 102: 874-880
6. Befus A D, Mowat C, Gilchrist M, et al. Neutrophil defensins induce histamine secretion from mast cells: mechanisms of action. J Immunol. 1999, 163: 947-953
7. Bensch KW, Raida M, Magert H J. hBD-1: a novel beta-defensin from human plasma. FEBS Letter. 1995, 368: 331
8. Bevins CL, Jones DE, Dutra A, Schaffzin J, Muenke M. Human enteric defensin genes: chromosome map position and a model for possible evolutionary relationships. Genomics. 1996, 31: 95-106
9. Boniotto M, Antcheva N, Zelezetsky I, Tossi A, Palumbo V, et al. A study of host defence peptide β-defensin 3 in primates. Biochemisty. 2003, 374: 707-714
10. Boniotto M, Tossi A, Del P, Sgubin, et al. Evolution of the β-defensin 2 gene in primates. Genes Immunity. 2003, 4: 251-257
11. Brierley, RA. Secretion of recombinant human insulin-like growth factor I (IGF-1). Methods Mol Biol. 1998, 103: 149-177
12. Bujacz G. Structure of monellin refined to 2.3 a resolution in the orthorhombic crystal form. Acta Crystallogr D Biol Crystallogr. 1997, 53(Pt 6): 713-9.
13.Cai S J(蔡绍晖),Du J(杜军),Chen X N(陈新年)et al.Recombinant human β-denfensin-2 gene expression in transfected COS-7 cell. Journal of West China University of Medical Sciences(华西医大学报).2000,31(4):433—437
14.Cai S J(蔡绍晖),Du J(杜军),Chen X N(陈新年),et al.Expression of recombinant human β-defensin-2 gene with C terminal of double Marks of Myc and poly-histidine in transfected COS-7 cells(C端带Myc和多聚组氨酸双标记的重组人β-防御素-2在COS-7细胞的转染表达).J Biomed Eng(生物医学工程杂志).2003,20(2):255-259
15.Cai S J(蔡绍晖),Huang N(黄宁),Wang B Y(王伯瑶).重组人β-防御素-2基因在昆虫细胞中的转染表达.Chinese joumal of pathophysiology(中国病理生理杂志).2001,17(8):800
16.Cai SH H(蔡绍晖),et al.Construction of plant expressive vector of human β-defensin-2 gene(重组人β-防御素-2基因的植物表达载体的建立).J Sichuan Univ(Med Sci Edi)(四川大学学报(医学版).2003.34(3):385-389
17. Caldwell JE, Abildgaard F, Dzakula Z, et al. Solution structure of the thermostable-sweet-tasting protein brazzein. Nat Stru Biol. 1998, 5-6: 427-431
18. Callewaert N, Larey W, Cadirgi H, et al. Use of HDEL-tagged trichoderma reesei mannosyl oligosaceharide 1,2-alpha-D-mannosidase for N-glycan engineering in Pichia Pastoris. FEBS letter. 2001, 503: 173-178
19. Cereghino Joan Lin, Cregg James M. Heterologous protein express in the methylotrophic yeast Pichia Pastoris. FEMS Microbiology. 2000, 24, 45-66
20. Chan B, Park H S, Kim SC. Mechanism of adion of antimicrobial peptide buforin Ⅱ: buforin Ⅱ kills microorganism by penetrating the all membrance and inhibiting celluar functions. Biochemical and biophysical Research Commucation. 1998, 244: 253-257
21.Chen shan(陈姗),He Fengtian(何凤田),et al.The cloning, high level expression in E. coli of human beta defensin 3 and its antimicrobial activity analysis. Chinese Journal of biotechnology(生物工程学报).2004,20(4):490—495
22.Chen Y(陈颖),Ge Y Q(葛毅强),Zhang L M(张利明),et al.Adyances in the research and Applicatin of mammal defensin,(哺乳动物防御素的研究进展及其应用前景).Prog Biochem Biophys(生物化学与生物物理进展).2001,28(1):17—21
23. Chertov O, Miciel DF, Xu L, Wang JM, et al. Identification of defensin-1, defensin-2, and CAP37/azurocidin as T-cell chemoattractant proteins released from interleukin-8-stimulated neutrophiis. J Biol Chem. 1996, 271, 2935-2940.
24.Chen Yumei(陈玉梅).酿酒酵母工程菌研究进展.生物工程进展.1993,13(1),11-16
25. Chertov O, Michiel D F, Xu L, et al. Identification of defensin-1,defensin-2 and CAP37/azurocidin as T cell chemoattractant proteins released from interleukin-8-stimulated neutrophils. J Biol Chem. 1996, 271: 2935-2940
26. Chung WO, Dale BA. Innate immune response of oral and foreskin keratinoeytes: utilization of different signaling pathways by various bacterial specirs. Infection and immunity. 2004, 1: 352-358
27. Cipalova I, Hostinova E, Gasperik J, Velebny V. High-level espression and purification of recombinant hBD-1 fused to LMN protein in E.coli. Protein Expr Purif. 2004, 37 (1): 207-212
28. Clare J J, Romanos M A, Rayment F B, et al. Production of mouse epidermal growth factor in yeast: high-level secretion using pichia pastries strains containing multiple gene copies. Gene. 1991, 105: 205-212
29. Clare JJ, Rayment FB, Ballantine SP, Screekrishna K, Romanos MA. High-level expression of tetanus toxin fragment C in pichia pastries strains containing multiple tandem integrations of the gene. Biotechnology. 1991, 9: 455-460
30.Cui H-ZH(崔洪志),Li M(李敏),Xu Q-F(徐琼芳),Guo S-D(郭三堆).Synthesis of a Bacterial-like Plant Monellin Gene and its Exptession in E. coli, Scientia Agriculture Sinica (中国农业科学).1999,32(1):58-62
31. Cociancich S, Ghazi A, Hetru C, Hoffmann J A, et al. Insect defensin, an inducible antibacterial peptide, forms voltage-dependent channels in Micrococcus luteus. J Biol Chem. 1993, 268: 19239-19245
32. Cole AM, Dewan P, Ganz T. Innate antimicrobial activity of nasal secrections. Infect Immun. 1999, 67: 3267-3275
33. Cregg JM, Madden KR, Barringer KJ. Function characterization of the two alcohol oxidase genes from the yeast Pichia Pastoris. Mol Cell Biol. 1989, 9: 1316-1323
34. Cregg JM, Vedvick T, Raschke W C, Recent advance in the expression of foreign genes in Pichia Pastoris. Biotechnology. 1993, 11: 905-910
35. Daniell S, Mellits K H, et al. Refolding the sweet-tasting protein thaumatin Ⅱ from insoluble bodies synthesized in E.coli. Food Chemistry. 2000, 71: 105-110
36. Daker K A, Lehrer R I, Ganz T. Isolation and characterization of human defensin cDNA clone. Pro Natl Acad Sci USA. 1989, 86(1): 342
37. Del P, Bonitto M, Zuccon D, Cervella P, Spano A, Amoroso A, Crovella S. β-defensin 1 gene variability among non-human primates. Immunogenetics. 2002, 53: 907-913
38. Diamond D L, Kimball JR, Krisanaprakomkit S, et al. Detection of β-defensins secreted by human oral epithelial cells. Journal of immunological methods. 2001, 256: 65-76
39. Diamond G, Zasloff M, Eck H, et al. Tracheal antimicrobial peptide, a novel Cysteine-rich peptide from mammalian tracheal nucosa: peptide isolation and cloning of cDNA. Pro Natl Acad Sci USA. 1991, 88(10): 3952-3956
40.Ding M(丁鸣),Hellekant G,Hu Zhong(胡忠).Characterization and chemicalmodifization of brazzein, a high potency thermostable sweet protein from pentadiplander brazzeana(耐热甜味蛋白Brazzein的特性和化学修饰).acta Botanica-Yunnanica(云南植物研究).1996,18(2):123～133
41. Donovan RS, Robinson CW, Glick BR. Review: optimizing inducer and cultrue condictions for expression of foreign proteins under the control of the Lac promoter. Journal of Industrial Microbiology. 1996, 16: 145-154
42. Duits L A, Rademaker M, Ravensbergen B, et al. Inhibition of hBD3, but not hBD-1 and hBD-2, mRNA expression by corticosteroids. Biochem Biophys Res Commun. 2001, 280(2): 522—555
43. Edens L, Bom I, Adrianus M, Ledeboer. synthesis and processing of the plant protein Thaumatin in Yeast. Cell. 1984, 37: 629-633
44. Edens L, Heslinga L, Klok R, et al. cloning of cDNA encoding the sweet tasting plant protein thaumatin and its expression in Escherichia coil Gone. 1982, 18: 1-2
45. Epand R M, Vogel H J. Diversity of antimicrobial peptides and their mechanisms of action. Biochim Biophys Acta. 1999, 1462: 11-28
46.Fan Changsheng(范长胜),甜蛋白的开发与应用研究.食品与发酵工业.2001,27(12):50-54
47. Fang X, Peng L, Xu Z, Wu J, Cen P. Cloning and expression of human beta-defensin-2 gene in Escherichia coil Protein Peptide Letter. 2002, 9(1): 31-7
48. Fariba M, Assadi-Porter, David J, Aceti, et al. sweetness determinant site of Brazzein, a small, heat-stable, sweet-tasting protein. Archieves of Biochemistry and Biophysics. 2000, 376(2): 259-265
49. Fariba M, Assadi-Porter, David J, Aceti, et al. efficient production of recombinant Brazzein, a small, heat-stable, sweet-tasting protein of plant orgin. Archieves of Biochemistry and Biophysics. 2000, 376(2): 252-258
50. Fans I. recent developments in the characterization and biotechnological production of sweeting-tasting proteins. Appllied Microbiological Biotechnology. 2000, 53: 145-151
51. Faus I, Patino Cristina, Del RJL, et al. expression of a synthetic gene encoding the sweet-tasting protein thaumatin in Escherichia coil Biochem Biophysi Resear Commu. 1996, 229: 121-127
52. Faus I, Del Moral C, Adroer N. secretion of the swotting-tasting protein thaumatin by recombinant strains of Aspergillus niger va Awamori. Appl. Microbiol Biotechnol. 1998, 49: 393-398
53. Faus I, Patino C, Del Rio J L, et al. expression of a synthetic encoding the sweet-tasting protein thaumatin in the filamentous fungus Penicillium roquewfortii. Biotechnology Letter. 1997, 19: 1185-1191
54. Frohlicho Poc, Young L G. Organization of human gene encoding the epididymis-specific EP2 protein variants and its relationship to defensin genes. Biol Report. 2001, 64: 1072-1079
55. Frye M, Bargon J, Gropp R. Expression of human beta defensin-1 promotes differentiation of keratinoeytes. J. Mol. Med. 2001, 79(5-6): 275-282
56. Ganz T, Selsted M E, Szklarek D, et al. Defensins, Natural peptide antibiotics of human neutrophils. The Journal of Clinical Investigation. 1985 76(4): 1427-35
57. Ganz T, Lehrer R I. Defensins, Current Opinion in Immunology. 1994, 6: 584-589
58. Gao Guang-Hua, Dia J X, Ming D, et al. solution conformation of brazzein by ~1H nuclear magnetic resonance: resonance assignment and secondary structure. International Journal of Biological Macromolecules. 1999, 24: 351-359
59.Gao SH N(高世年)主编.《实用食品添加剂》.天津科学技术出版社.2000,197-207
60. Garcia J R, Jaumann F, Schultz S, Krause A, et al. Identification of a noval, multifunctional β defensin (human β defensin 3 )with specific antimierobial activity. Cell tissue Res. 2001, 306: 257-264
61. Garcia J R, Krause A, Schulz S, et al. Human β-defensin-4: a novel inducible peptide with a special salt-sensitive spectrum of antimicrobial activity. The FASEB Journl. 2001, 15: 1819-1821
62. Garcia JR, Jaumann F, Schulz S, et al. Identification of a noval, multifunctional beta-defensin(human beta-defensin 3) with special antimicrobial activity, its interaction with plasma membrance of Xenopus oocytes and the induction of macrophgge chemoattraction. Cell Tissue Res. 2001, 306(2): 257-264
63. Goochee C F, Gramer M J, Andersen D C, Bahr J B, Rasmussen J R. The oligosaccharides of glycoproteins: bioprocess factors affecting oligosaccharide structure and their effect on glycoprotein proteins. Biotechnology. 1991, 9: 1347-1355
64.Guo L D(郭良栋),Qian Sh J(钱世钧),Ye J(叶军),Jiao Q H(矫庆华).Studies on the culture conditions of engineered strain of human lysozyme(人溶菌酶工程菌培养条件的研究).Acta Microbiologica Sinica(微生物学报).1997,37(1):53-57
65.Guo M J(郭美锦),Zhumag Y P(庄英萍),et al.Expression of recomibinant human serum albumin in genetically enginnred pichia pastoris in high-density fermentation(重组巴氏毕赤酵母高密度发酵表达rHSA).Acta Microbiologica Sinica(微生物学报).2002,42(1):62-68
66. Hancock REW, Falla T and Brown MH. Cationic bacteriecidal peptides. Adv Microb Physiol. 1995, 37: 135-175.
67. Hancock Robert EW. Cationic peptides: effectors in innate imminity and novel antimicrobials, the Lancet Infectious Dieases. 2001, 11(1): 156-164
68. Hancock REW, Chapple D S. peptide antibiotics. Antimicrob Agents Chemother. 1999, 43: 1317-1323
69. Harder J, Bartels J, Christophers E, Schroder JM. A peptide antibiotic from human skin. Nature. 1997, 387 :861
70. Harder J, Bartels J, Christophers E, Schroder JM. Isolation and characterization of human β-defensin-3, a novel human inducible peptide antibiotic. Journal of Biological Chemisity. 2001, 276(8): 5707-5713
71. Hasslacher M, et al. Hight-level intracellular expression of hydroxynitrile lyase from the tropical rubber tree hevea brasiliensis in microbial hosts. Pro Expr Purif. 1997, 11, 61-71
72. Haynes R J, Tighe P J, Dua H S. Antimicrobial defensin peptides of the human ocular surface. Br f Ophthalmol. 1999, 83: 737-741
73.He Cheng(何诚),Zhu Yunsong(朱运松).甲醇营养型酵母表达系统的研究进展.生物工程进展.1998,18(3),7-11
74. Hoffmann JA, Hertu C. Insect defensins: inducible antibacterial peptides. Immunol Today. 1992, 13(10): 411-415
75. Hoover D, Wu Zhibin, Tucker K, Lu W, Lubkowski J. Antimicrobial characterization of human β-defensin 3 derivatives. Antimicrobial agent and chemotherapy. 2003, 9: 2804-2809
76. Hoover D, Rajashankar K R, et al. the structure of human β-defensin-2 shows evidence of higher order oligimerization. The journal of biological chemistry. 2000, 275(42): 32911-32918
77.Hu Zhong(胡忠),He ming(何敏).study on Mabinlin, a sweet protein from the seeds of capparis masaikai levi Ⅰ extraction, purification and certain characteristics(吗槟榔甜蛋白的研究Ⅰ提取、纯化和某些特性).Acta Botanica yunnanica(云南植物研究).1983,5(2):207-212
78.Huang Yadong(黄亚东),Zheng Qi(郑青),et al.Modification of Cecropin AD gene and its expression in Pichia pastoris. Journal of China university of technology(Natural science edition)(华南理工大学学报,自然科学版).2002,30(2):13—16
79.Huang Yin(黄音),Liu Feipeng(刘飞鹏),Zhou Tianhong(周天鸿),Zhu Jiaming(朱嘉明).蛙皮素—蜂毒素杂合肽基因在大肠杆菌的克隆与诱导表达.生物工程学报.2001,17(2):207-210
80. Illingworth C, Larson G Hellenkant G secretion of the sweet-tasting protein Thaumatin by Bacillus Subtilis. Biotechnology Letter. 1988, 16: 587-592
81. Ishkawa K, Ota M, Ariyoshi Y, Sasaki H, et al. Crystallization and preliminary X-ray analysis of brazzein, a new sweet protein. Acta Crystallogr D. 1996, 52: 577-578
82. Izawa H, Ota M, Kohmura M, Ariyoshi Y. Synthesis and characterization of the sweet protein brazzein. Biopolymers. 1996, 39(1): 95-101
83. Jia H P, Schutte B C, Schudy A, et al. Discovery of new human beta-defensins using a genomics-based approad. Gene. 2001, 263: 211-218
84.Jiang Y(江阳),Shi H Zh(时宏珍),et al.Study on the expression of recombination of human Interleukin-13(rhIL-13) in methylotropic yeast pichia pastoris and its effect on the induction of DC to differentiate in vitro.中国免疫学杂志.2003,19:82—87
85. Jin Z, Danilova V, Assadi-Porter FM, Aceti DJ, Markley JL, Hellekant G Critical regions for the sweetness of brazzein. FEBS Lett. 2003, 544(1-3): 33-7
86. Jin Z, Danilova V, Assadi-Porter FM, Markley JL, Hellekant G. Monkey electrophysiological and human psychophysical responses to mutants of the sweet protein brazzein: delineating brazzein sweetness. Chem Senses. 2003, 28(6): 491-8
87. Joly S, Maze C, Mccray P B, Guthmiller J M. Human β-defensin 2 and 3 demonstrate strain-selective activity against oral microorganisms. J Clin Microbiol. 2004, 3: 1024-1029
88. Jones DE, Bevins CL. Paneth cell of the human small intestine express an antimicrobial peptide gene. J boil Chem. 1992, 267: 23216-23215
89. Jones DE, Bevins CL. Defensin-6 mRNA in human Paneth cell: implications for antimicrobial peptides in hose defensin of the human bowel. FEBS Letter. 1993, 315: 187-192
90. Kapralek F, Jecmen P, Sedlacek J, Fabry M, Zadrazil S. Fermentation conditions for high level expression of tac-promoter-controlled calf prochymosin cDNA in E.coli HB101. Biotechnology and bioengibeering. 1991, 37: 71-79
91. Keneth JM, Roert IL. β-sheet antibiotic peptic as potential detal therapeutics. International Journal of Antimicrobial Agents. 1998, 9: 269-280
92. Kim SH, Kang CH, et al. redesingniting a sweet protein: increased stability and renaturability. Protein Engineering. 1989, 2(8): 571-579
93. Kohmura M, Nio N, Ariyoshi Y. Highly probable active site of the sweet protein monellin. Biosci Biotechnol Biochem. 1992, 56(12): 1937-42
94. Kondo K, Miura Y, Sone H, et al. high-level expression of a sweet protein monoeilin, in the food yeast Candida utilis. Nat Biotechnol. 1997, 15: 453-457
95.Kong J Q(孔建强),Zhao Q(赵琦),Gao Y(高音),et al.Sweet protein thaumatin and it's genetic engineering(奇异果甜蛋白及其基因工程).Hereditas(beijing)(遗传).2003,25(2):232—236
96. Koutz P, Davis GR, Stillman C, Barringer K, et al. Structural comparison of the Pichia Pastoris alcohol oxidase genes. Yeast. 1989, 5: 167-177
97. Krisanaprakornkit S, Weinberg A, Perez CN, et al. Expression of the peptide antibiotic human beta-defensin 1 in cultured gingival epithelial cell and gingival tissue. Infection and immunity. 1998, 66: 4222-4228
98. Lee SY, Lee JH, Chang HJ, Cho JM, Jung JW, Lee W. Solution structure of a sweet protein single-chain monellin determined by nuclear magnetic resonance and dynamical simulated annealing calculations. Biochemistry. 1999, 38(8): 2340-6.
99. Lehrer RI, Barton A, Dither KA. Interaction of human defensins with E.coli mechanism of bactericidal activity. J Ciin Invest. 1989, 84(2): 553-561
100. Lehrer RI. defensin: antimicrobial and cytotoxic peptides of mammalian cells. A.Rev Immun. 1993, 11: 105-128
101. Lehrer RI, Ganz T. Antimicrobial peptides in mammalian and insect host defence. Cur opin Immu. 1999, 11: 23-27
102. Lehrer RI, Ganz T. Endogenous vertebrate antibiotics defensins, protegrins and other cysteine-rich antimicrobial peptides. Ann NY Acad Sci. 1996, 797: 228-239
103.Li Guangkuo(李广阔),Li X(李旭),Tian F Zh(田伏洲),et al.人源性空回肠粘膜防御素-5的基因的克隆.中华实验外科杂志.2003,20(10):884-885
104.Li Hongiun(李洪军).防御素研究进展.生物工程进展.2001,21(3):34-38
105.Li Hongzhao(李洪钊),Li Liangzhu(李亮助),Sun Qiangming(孙强明),et al.Strategies for optimizing pichia pastoris expression syetem(巴斯德毕氏酵母表达系统优化策略).Acta Microbiologica sinica(微生物学报).2003,43(2):288-292
106.Li G L(李广林),Guo A G(郭蔼光),Fan S H(范三红).Cloning and expression of brazzein gene(植物Brazzein基因的克隆与表达).Acta Bol Boreal-Occident Sin(西北植物学报).2004,24(7):1271-1274
107.Li Jing(李晶),Zhao Xiaoxiang(赵晓样).甲醇酵母基因表达系统的研究进展.生物工程进展.1999,19(2),17-20
108.Li X D(李晓东),Liu J M(刘敬梅),Chen H(陈杭),et al.Genetic transformation of sweet protein MBLⅡ on tomato(甜蛋白基因MBLⅡ对番茄的遗传转化).Acta Bol Boreal-Occident Sin(西北植物学报).2004,24(5):808-811
109.Li Y(李英),Yin J(尹俊),Zhang Sh J(张树军).Synthesis and cloning of cDNA encoding the sweet plant protein monellin(植物甜蛋白monellin基因的合成和克隆).Journal of Inner Mongolia Agricultural University(内蒙古农业大学学报).2001,22(3):35-38
110. Liao YF, Lal A, Moremen KW. Cloning, expression, purification and characterization of the human broad specificity lysosomal acid alpha-mammosodase. Journal of biology and chemistry. 1996, 271, 28348-28358
111. Lichtenstein A. Mechanism of mammalian cell lysis mediated by peptide defensins, Evidence for an initial alternation of the plasma membrance. J Clin Invest. 1999, 88(1): 93-100
112.Ling GD(凌关庭)主编.《食品添加剂手册》(第三版).化学工业出版社.2003,645-666
113. Linzmeier R, Miehaelson D, Liu L, et al. The structure of neutrophil defensin genes. FEBS Letter. 1993, 321: 267-273
114. Linzmeier R, Ho C H, Hoang BV, Ganz T. a 450-kb of defensin genes on human chromosome 8P23. gene. 1999, 233: 205-211
115.Liu J M(刘敬梅),Chen D M(陈大明),Chen H(陈杭).Genetic transformation and plant regeneration of lettuce with sweet protein gene MBLⅡ(甜蛋白基因MBLⅡ对莴苣的遗传转化).Acta Horticulturae Sinica(园艺学报).2001,28(3):246-250
116. Liu L, Wang L, Jia H P, et al. Structure of the mapping of the human β-defensin HBD-2 gene and its expression at sites of inflammation. Gene. 1998, 222: 237-244
117.Liu Q Y(刘秋云),He ZH M(贺竹梅),Cao J(曹俊),et al.利用农杆菌系统将超甜蛋白基因导入烟草.遗传.1998,20(1):24-27
118. Liu W C, Zhang XY, Hu JL, tumor cell eytolysis by defensins of human neutrphils in vitro. Inter J Modern cancer ther. 1998, 1: 45
119.Liu W C(刘文超),Mu H X(穆怀兴),Ren J(任军),Zhang X Y(张学庸),et al.防御素对胃癌细胞系体外的杀伤作用.World Chin J Digestol(世界华人消化杂志).2001,9(6):622-626
120. Liu X, Hu Z, Maeda S, Aiuchi T, Nakaya K. Purification, complete amina acid sequence and structure chaeterization of the heat stable sweet protein, mabinlin Ⅱ. Eur J Bioehem. 1993, 211: 281-287
121.Lu B(陆兵),Xia J(夏杰),Xu D Sh(徐殿胜),Chen G H(陈国豪).The Lactose induction for the expression of rhNTA under the control of φ10 promoter in E. coli(φ10启动子控制下的大肠杆菌中rhNTA表达的乳糖诱导).Journal of East China university of science and technology(华东理工大学学报).2001,4:131-134
122.Ma li(马骊),Wang Xiaoning(王小宁),Zhang Zhiqing(张智清),et al.Expression, Purification and biological activity analysis of human vascular endothelial growth factor (VEGF_(165)) in pichia pastoris(人血管内皮因子(VEGF_(165))在毕氏酵母中的表达及生物活 性分析).Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(3):325-330
123. Mackewicz C E, Yuan J, Tran P, et al. alpha-defensins can have anti-HIV activity but are not CD8 cell anti-HIV factors. AIDS. 2003, 17(14): 31
124. Malsetta G, Batoni G, Esin S, et al. Activity of Human β-defensins 3 alone or comibined with other antimicrobial agent against oral bacteria. Antimicrobial agent and chemotherapy. 2003, 10, 3349-3351
125. Makrides SC, strategies for achieving high-level expression of genes in E.coli. Microbiological review. 1996, 60(3): 512-516
126. Mallow EB, Harris A, Salzman N, et al. Human enteric defensins. gene struction and developmental expression. J Biol Chem. 1996, 271 (8): 4038-4045
127. Masuda T, Tamaki S, Kaneko R, Wada R, Fujita Y, Mehta A, Kitabatake N. Cloning, expression and characterization of recombinant sweet-protein thaumatin Ⅱ using the methylotrophic yeast Pichia pastoris. Biotechnoi Bioeng. 2004, 85(7): 761-9.
128. Masuda J A, Nirasawa S, Nakaya K, Kurihara Y. Cloning and sequencing of a cDNA encoding a taste-modifying protein miraculin. Gene. 1995, 161: 175-177
129. Mathews M, Jia H P, Guthmiller J M, et al. Production of beta-defensin antimicrobial peptides by oral mucosa and salivary gland. Infection and Immunity. 1999, 67: 2740-2745
130. Mineshiba F, Takashiba S, Mineshiba J, Matsuura K, Kokeguehi S, et al. Antibacterial activity of synthetic human β-denfensin 2 against periodontal bacteria. J Int Acad Periodontal. 2003, 5: 35-40
131. Ming D, Hellekant G, Brazzein. a new high potency sweet protein from Pentadiplandra brazzeana Baillon. FEBS Letter. 1994, 355: 106-108
132. Moralejo FJ, Cardoza RE, Gutierrz S, et al. Thaumtin production in Aspergillus awamori by use of expression cassettes with strong fungal promoters and high gene dosage. Applied and Environmental Microbiology. 1999, 65 (3): 1168-1174
133. Moralejo F J, Cardoza R E, Gutierrz S, et al. overexpression and lack of degradation of thaumatin in an aspergillopepsin A-defective mutant of Aspergillus awamori containing an insertion in the pepA gene. Appl Microbiol biotechnol. 2000, 54: 772-777
134. Morris J A, Cagan R H. purification of Monellin, the sweet principle of Discoreophyllum curarainisii. Biochem. Biophys Acta. 1972, 261: 114-122
135. Muller C, Markovie-Lipkovski J, Klatt T. human α-defensins HNPs-1,-2,and-3 in renal cell carcinoma influences on tumor cell proliferation. American journal of patbology. 2002, 160(4): 1311-1323
136. Nirasawa S, Nishino T, Katahira M, et al. Struction of heat-stable and unstable homologues of the sweet protein mabinlin. Eur J Biochem. 1994, 223: 989-995
137. Ogawa M, Nakamura S, Atsuchi T, et al. Macromolecu lar antimicrobial glycoprotein, achacin, expressed in a methylotrophic yeast Pichia Pastoris. FEBS Letters. 1999, 448: 41-44
138. Ohtani W, Nawa Y, Takeshima K, Kamuro H, kobayashi K, Ohmura T. Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia Pastori. Anal Biochem. 1998, 256, 56-62
139. Ortega M R, Ganz T, Milner S M. Human beta defensin is absent in bum blister fluid. Burns. 2000, 26: 724-726
140. Periathamby AR, Andrew RD. Current status of defensins and their role in innate and adaptive immunity. FEMS Microbiology Letters. 2002, 206: 9-18
141. Pang SK, Oberhans SM, Rasmussen J K, et al. Expression of gene encoding a scorpion insectotoxin peptide in yeast, bacteria and plants. Gene. 1992, 116: 165-172
142. Penarrubia L, Kim R, Giovannoni J. production of the sweet protein monellin in transgenic plants. Biotechnol. 1992, 10: 561-564
143. Peng L, Xu Z, Fang X, Wang F, Yang S, Cen P. Preferential codons enhancing the expression level of human beta-defensin-2 in recombinant Escherichia Coli. Protein Pept Lett. 2004, 11(4): 339-344
144. Peng Yu-Cai, et al. Production of active polyomavirus large T antigen inyeast Pichia Pastoris. Virus Reasearch. 1997, 49: 41-47
145.Peng Li(彭力),Xu Zhinan(徐志南),et al.Advances in the research of human defensins. Chinese Journal of biotechnology(生物工程学报).2003,19(3):261-266
146.Peng Yi(彭毅),Bu Wei(步威).甲醇酵母表达系统.Biotechnology Information(生物技术通报).2000,1,38-41
147.Peng Yi(彭毅),Yang Xicai(杨希才),Kang liangyi(康良仪).影响甲醇酵母中外源蛋白表达的因素.Biotechnology Information(生物技术通报).2000,4:33-36
148. Peschel A, Ralph W. Jack, Michael Otto. Staphylococcus aureus Resistance to Human Defensins and Evasion of Neutrophil Killing via the Novel Virulence Factor MprF Is Based on Modification of Membrane Lipids with L-Lysine. The Journal of Experimental Medicine. 2001, 193(9): 1067-1076
149. Piers K L, Brown M H, Hancock REW. Recombinant DNA procedures for producing small antimicrobial cationic peptide in bacteria. Gene. 1993, 134(1): 7-13
150. Prohaszka Z, Nemet K, Csermly P, et al. defensins purified from human granuloeytes bind C1q and activate the classical complement pathway like the transmembrane glyeoprotein gp41 of HIV-1. Molecular immunology. 1997, 34(11): 809-816
151.Qu X D(曲晓丹),Wang Ai X(王爱霞).人中性粒细胞防御素的提纯及活性分析.中华医学杂志.1991,71(11):616—619
152. Quinones-mateu M, Lederman M M, Feng Z, et al. Human epithelial-defensin 2 and 3 inhibit HIV-1 replication. AIDS. 2003, 17(16): 39-43
153. Raj PA, Antonyraj KJ, Karunakaran T. Large-scale synthsis and functional elements for the antimicrobial activity of defensins. Biochem J. 2000, 347: 633-64
154. Reiehart J M, Petit I, legtain M, et al. expression an inducible antibacterial peptide from the fleshly Phormia terranovae. Invert reproduct develop. 1992, 21 (1): 15-24
155.Ren H Y(任洪艳),Zhang L H(张苓花),Wang Y J(王运吉).Cloning of human α-defensin 1 and iu structural analysis(人α-防御素HNP-1基因克隆及其结构分析).Journal of Dalian institute of light industry(大连轻工学院学报).2003,22(4):265-269
156. Risso A. Leukocyte antimicrobial peptide: multifunetional effector molecules of innate immunity. J Leukocyte Biol. 2000, 68: 785-792
157. Robert IL, Tomas Ganz. Defensins of vertbrate animal. Current Opinion in Immunology. 2002, 14: 96-102
158. Romanos MA, Clare JJ, Beesley KM, Rayment FB. Recombinant Bordetella Pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties. Vaccine. 1991, 9: 901-906
159. Romanos Mike, Wellcome Glaxo, Beckenham. Advances in the use of pichia pastoris of high level gene expression. Current Opinion in Biotechnology. 1995, 6: 527-533
160. Sahly H, Schubert S, Harder J, et al. Bukholderia is highly resistant to human beta-defensin 3. Antimierobial agents and chemotherapy. 2003, 5: 1739-1741
161. Sambrook Joseph, Russell David W. Molecular Cloning: A laboratory Manual. Cold Spring Harbor Laboratory Press. 2001
162. Sehroder JM. Epithelial peptide antibiotic. Biochem Pharma. 1999, 57: 121-134.
163. Sehroder JM, Harder J. Molecules in focus human beta-defensin-2. The international journal of biochemistry & cell biology. 1999, 31: 635-651
164. Scorer CA, Clare JJ, McCombin WR, et al. Rapid selection using G418 of high copy number transformants of Pichia Pastoris for high-level foreign gene expression. Bioteehnoiogy. 1994, 12: 181-184
165. Selsted ME. Primary struction of three human neutrophlis defensins. The Journal of Clinical Investigation. 1985, 76: 1436-40
166. Selsted M E, Tang Y Q, Morris W L, Mcguire P A, et al. Purification primary structures and antibacterial activities of β-defensins, a new family of antimicrobial peptides from bovine neutrphils. J Biol Chem. 1993, 268: 6641-6648
167. Seo SJ, Ahn SW, Hong CK, Ro Bl. Expression of β-defensins in human kerationoeyte cell lines, Journal of dermatological science. 2001, 27: 183-191
168.Shao L L(邵玲莉),Zhou N D(周楠迪),Jin J(金坚),et al.Optimization of culture medium for the production of recombinant human angiostatin(重组人血管抑制素生产菌培养基优化研究).Pharmaceutical biotechnology(药物生物技术).2004,11(2):91—95
169.Shao L L(邵玲莉),Zhou N D(周楠迪),Wu Q F(吴奇凡),Jin J(金坚),et al.Fermentation contion of angiostatin by recombinant(重组大肠杆菌生产人血管抑制素的发酵条件).journal of Wuxi university of light industry(无锡轻工大学学报).2004,23(2):8—11
170. Sharma S, Verma I, Khuller GK. Therapeutic potential of human neutrophil peptide 1 against experimental tuerculosis. Antimicrobial agent and chemotherapy. 2001, 2: 639-640
171. Sharp PM, Tuohy TMF, Mosurski KR. Codon usage in yeast: cluster analysis clearly differentiates highly and lowly expressed genes. Nuleid acid Research. 1986, 14(13): 5215-5143
172. Shen S, Sulter G, Jeffreies T W, et al. A strong nitrogen source-regulated promoter for controlled expression of foreign genes in the yeast pichia pastoris. Gene. 1998, 216: 93-106
173.Shiqiang(石强),Liu Feipeng(刘飞鹏).抗菌肽基因的表达和转基因研究现状.生物工程进展.2000,20(11):37—40
174. Slootstra JW, De Geus P, Haas H, Verrips CT, Meloen RH. Possible active site of the sweet-tasting protein thaumatin. Chem Senses. 1995, 20(5): 535-43
175. Somoza JR, Cho JM, Kim SH. The taste-active regions of monellin, a potently sweet protein. Chem Senses. 1995, 20(1): 61-8.
176. Sreekrishna K, Brankamp RG, Clare JJ, et al. strategies for optimal synthesis and secretion of heterogous proteins in the methylotrophic yeast pichia pastoris. Gene. 1997, 190: 55-62
177. Spadaccini R, Crescenzi O, Tancredi T, De Casamassimi N, et al. Solution structure of a sweet protein: NMR study of MNEI, a single chain monellin. J Mol Biol. 2001, 305(3): 505-14
178. Suami T, Hough L, Machinami T, et al. Molecular mechanisms of sweet taste: The sweet protcin, Thaumatin I. Food chemistry. 1997, 60(3): 277-285
179.Tang Bing(唐彬),Huang Ning(黄宁),Wu Qi(吴琦),et al.Studies on the expression of transferred human α defensin HNP1 gene in tracheal eoithelial cells cultivated in vitro. Chinese Journal of Microbiology(中华微生物学和免疫学杂志).1999,19(6):481-484
180.Tuo xiaoye(庹晓晔),Xu Mingda(徐明达),Chen Bi(陈璧),et al.Fusion expression of human beta-defensin 3 in E.coli and primary analysis of its antimicrobial activities. Bull Acad Mil Sci(军事医学科学院院刊).2004,28(2):114-122
181. Urabatsch IL, Wilke MS, Gimi K, et al. Purification and characterization of N-glycosylation mutant mouse and human P-glycoproteins expressed in pichia pastoris cells. Arch Biochem Biophys. 2001, 388: 171-177
182. Valore EV, Park C H, Quayle A J, et al. Human beta-defensin-1: an antimicrobiai peptide of urgenital tissues. Journal of clinical investigation. 1998, 101 (8): 1633-1642
183. Van del wel H. Structure modification of sweet proteins and its influence on sensory properties. Chemistry and Industry. 1983, 3: 19
184. Van del wel H, lyengar RB. Assignment of the disulfide bonds in the sweet tasting protein thaumatin I. Eur J Biochem. 1984, 144: 41
185. Van der Wel H, Larson G. Isolation and charcterization ofpentadin, the sweet principle of Pentadiplandra brazzeana Baillon. Chem Senses. 1989, 14: 73-79
186. Van der Wel H, Loeve K. Isolation and charcterization of Thaumatin Ⅰ and Ⅱ, the sweet tasting protein from Thaumatotoccus daniellii Benth. Eur J Biochem. 1972, 31: 221-225
187. Valore EV, Park CH, Quayle AJ, et al. Human beta-defensin 1: an antimierobial peptide of urogenital tissue. Journal of clinical investigation. 1998, 101(8): 1633-1642
188. Wada A, Ogushi K, Kimura T, et al. Helicobaeter pylori-mediated transcriptional regulation of the human beta-defensin 2 gene requires NF-kappaB. Cell Mierobiol. 2001, 3 (2): 115-123
189. Wade D, Boman A, Waklin. amino acid-containing channel-forming antibiotic peptides. Proc.Natl Acad Sci USA. 1990, 87: 4761-4765
190. Wang F, Fang X, Xu Z, Peng L, Cen P. Fusion expression of human beta-defensin-2 from multiple joined genes in Escheriehia coli. Prep biochem Bioteehnol. 2004, 34 (3): 215-225
191.Wang Tongying(王同映),Wang Changmei(王昌梅),Wei Guang(魏光),et al.Expression of the recombinant human Intedekin-Ⅱ in pichia pastoris(重组人白细胞介素Ⅱ在毕氏酵母中的表达).Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(6):659-664
192.Wang Weirong(王维荣),Liu Aiming(刘爱民),Qian Zhikang(钱志康),Huang Weida (黄伟达).青岛海葵强心活性多肽在毕氏酵母中的分泌表达.生物化学与生物物理进展.2000.27(4):407-411
193.Wang Yanru(王艳茹),Ren Hongwei(任宏伟),et al.人三叶因子3在毕赤酵母中的表达及活性分析.生物化学与生物物理学报.2001,33(6):653-658
194. Waterham HR, Digan ME, et al. Isolation of the pichia pastoris glyceraldehydes-3-phosphate dehydrogenase gene and regulation and use of its promoter. Gene. 1997, 186: 37-44
195. Weichert MJ, Pagratis M, Curry SR, Blackmore R. stabilization of apoglobin by low temperature increases yield of soluble recombinant hemoglobin in E.coli. Applied and environmental microbiology. 1997, 11: 4313-4320
196. Wimley WC, Seleted ME, White SH. Interactions between human defensins and lipid bilayers: evidence for formation of multimeric pores. Protein Science. 1994, 3: 1362-1373
197. Withers-Martinez C, carpenter EP, Hackett F, et al. PCR-based gene synthesis as an efficient approach for expression of the A+T-rich malaria genome, protein engineer. 1999, 12: 113-1120
198. Witty M. propeothaumatin Ⅱ is processed to biological activity in solanum tuberosum. Biotechnol Letter. 1990, 12: 131-136
199.Wu G(吴刚),Wang Y M(汪亚平),Zhu Z Y(朱作言).Secretion expression of Monellin in Pichia Pastoris(甜蛋白Monellin在毕赤酵母中的分泌表达).高技术通讯.2003,11:20-23
200. Wu M, Maier E, Bena R, Hancock REW. Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemstry. 1999, 38: 7235-7234
201.Xin Li(辛利),Zhang Li(张励),et al.人血管抑制素在毕赤酵母中的表达及活性测定.Acta biochemical and biophysics sinica(生物化学与生物物理学报).2001,33(3):291-295
202.Xiong Aisheng(熊爱生),Peng Rihe(彭日荷),Li Xian(李贤).Influence of signal peptide sequences on the expression of heterogeneous proteins in pichia pastoris(信号肽序列对毕氏酵母表达外源蛋白质的影响).Acta Biochimica et Biophysica Siniea(生物化学与生物物理学报).2003,35(2):154-160
203. Xiong L, Sun S. Molecular cloning and transgenic expression of sweet protein mabinlin in potato tubers. Plant Physiol. 1996, 111: 57
204.Xu ZH N(徐志南),Liu G(刘刚),Cen P L(岑沛霖).Research on fermentation for hEGF production in recombinant Escherichia coli(重组大肠杆菌生产hEGF发酵条件的研究).Journal of Zhejiang universiq4engineering science)(浙江大学学报(工学版)).2001,35(1):25-28
205.Yang Ch L(杨成丽),Liu Sh N(刘树楠),Zhou J Y(周吉源),et al.Introduction of Thaumatin gene into Tobacco(Nicotiana tabacum L)through Agrobacterium-mediated genetic transformation(高效烟草遗传转化体系的建立及其甜蛋白基因的导入).Biotechnology(生物技术).2004,14(2):9-11
206. Yang D, Biragyn A, Kwak L W. Oppenheim J J, Mammalian defensins in immunity: more than just microbicibal. Trends Immunol. 2002, 23: 291-296
207. Yang D, Chertov D, et al. beta-defensin: linking innate and adaptive immunity through dentritic and T cell CCR6. Science. 1999, 286: 525-528
208. Yang D, Chen Q, Chertov O, et al. Human neutrophil defensins selectively chemoattract naive T and immature dendritic cells. J leukoc Biol. 2000, 68: 9-14
209.Yang M ZH(杨美珠),Pan N S(潘乃楗),Chen ZH L(陈章良).高效马铃薯遗传转化体系的建立及甜蛋白基因的导入.植物学报.1992,34(1):31-36
210.Yang X J(杨晓娟),Chen J(陈静),Ma X R(马欣荣),et al.人β-防御素-2植物表达载体的构建.四川大学学报(自然科学版).2002,39:55-59
211.Yao Bing(姚斌),Zhang Chunyi(张春义),Wang jianghua(王建华),Fan Yunliu(范云六).高效表达具有生物活性的植酸梅的毕氏酵母.中国科学(C辑).1998,28(3):237—243
212.Ye J(叶娇),Chen Ch H(陈长华),Yang Y Q(杨雅琴),et al.expression of recombinant human superoxide dismutase gene in E.coli induced by Lactosc(乳糖诱导重组人SOD基因 在大肠杆菌中的表达).Chinese journal of pharmaceuticals(中国医药工业杂志).2001,32(2):536-539
213.Yu Xuehong(于学红),Yuan Hanying(袁汉英),et al.Expression of Monellin, a sweet protein, in pichia pastoris. Journal of Fudan University(Natuml science)(复旦学报 自然科学版).2002,41(6):650-654
214.Yuan Hanying(袁汉英),Ai HS(艾华水),Fan ZS(范长胜)et al.Construction of the expression vector for single-chain Monellin gene and its expression(单链莫热宁基因表达载体的构建及其分泌表达).Journal of Fudan University(Natural science)(复旦学报 自然科学版).2003,42(4):
215. Zhang L, Falla T, Wu M, et al. Determination of recombinant production of antimicrobial cationic peptides and creation of peptide variants in bacteria. Biochemical and Biophysical Research Commutation. 1998, 247: 674-680
216.Zhang Chunyi(张春义),Zhan jun(赵军),Wu Changjian(武长剑),Fan Yunliu(范云六).利用毕氏酵母表达抗真菌鳖素基因.科学通报.1998,43(19):2085-2089
217. Zhang G L, Wu H, Shi J. Molecular cloning and tissue expression of procine β-defensin-1. FEBS Letter. 1998, 424: 37-40
218.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).Expression of human defensin —1(HNP-1)in E.coli. Chinese biochemical journal(生物化学杂志).1997,13(3);264-269
219.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).human defensin 1(HNP-1) expression in Tobacco: A preliminary observation on anti-TMV activity of transgenic Tobacco plants. Acta Botaniea sinica(植物学报).1997,39(6):574-576
220.Zhang Manchao(张满朝),Zheng Guochang(郑国锠).Transient expression on human defensin 1(HNP-1)in Tobacco protoplast. Acla Biologiae experimentalis sinica(实验生物学报).1997,30(4):209-211
221.Zhang Yi(张毅),Qu X M(屈贤铭),Yang Sh L(杨胜利).High cell density culture of the recombinant E.coli harboring h GM-CSF/IL-3 fusion expression plasmid(重组GM-CSF/IL-3融合蛋白表达菌发酵条件的研究).Industrial Microbiology(工业微生物).2000,30(2):8-11
222.Zhao Donghong(赵东红),Dai Zhuying(戴祝英),Zhuo Kaiya(周开亚).昆虫抗菌肽的功能、作用机理与分子生物学研究最新进展.生物工程进展.1999,19(5):14—18
223. Zhao C Q, Wang I, Lehrer R I. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Letters. 1996, 396: 319-322
224.Zhou J H(周家华),Cui Y D(崔英德),Li B N(黎碧娜)等主编.《食品添加剂》.化学工业出版社.2001
225.Zhou P(周平),Pan N S(潘乃稽),et al.外源甜蛋白Thaumatin基因转入烟草的研究.植物学通报.1994,11(4):17-20