鹅ASC基因序列分析与组织表达差异研究
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摘要
炎症小体作为固有免疫的重要组分在机体免疫反应和疾病发生过程中具有重要作用,ASC基因是炎症小体中连接胞浆受体和Caspase-1的关键蛋白.为进一步了解鹅的炎症系统,参考NCBI上公布的预测序列(XM_013201308.1)设计引物,克隆了马岗鹅ASC基因(go-ASC),并进行了生物信息学分析和组织表达差异研究.结果显示,go-ASC基因完整ORF长663 bp,共编码220个氨基酸.同源性分析显示,go-ASC基因序列与哺乳类动物的同源性较低(41.0%～52.6%),结构域也存在较大差异,仅含有CARD结构域,缺乏PYD结构域.组织表达结果显示,go-ASC基因主要在血液和粘膜上皮组织表达,在肌肉组织中表达量最低.
As an important component of innate immunity, inflammatory bodies play an important role in the body's immune response and disease development. ASC is a key protein in the inflammatory body that connects cytoplasmic receptors and Caspase-1. In order to further understand the inflammatory system of goose, the primers were designed with reference to the predicted sequence report in NCBI(XM_013201308.1), and the ASC gene of Magang goose was cloned, and bioinformatics analysis and tissue expression difference were carried out. The results showed that the complete ORF region of the go-ASC gene was 663 bp in length and encoded 220 amino acids. Homology analysis showed that the go-ASC gene sequence has low homology with that in mammals(41.0%～52.6%), and the domain is also largely different. It only contains CARD domain and lacks PYD domain. The results of tissue expression showed that the go-ASC gene was mainly expressed in blood and mucosal epithelial tissues, and the expression level was the lowest in muscle tissue.
As an important component of innate immunity, inflammatory bodies play an important role in the body's immune response and disease development. ASC is a key protein in the inflammatory body that connects cytoplasmic receptors and Caspase-1. In order to further understand the inflammatory system of goose, the primers were designed with reference to the predicted sequence report in NCBI(XM_013201308.1), and the ASC gene of Magang goose was cloned, and bioinformatics analysis and tissue expression difference were carried out. The results showed that the complete ORF region of the go-ASC gene was 663 bp in length and encoded 220 amino acids. Homology analysis showed that the go-ASC gene sequence has low homology with that in mammals(41.0%～52.6%), and the domain is also largely different. It only contains CARD domain and lacks PYD domain. The results of tissue expression showed that the go-ASC gene was mainly expressed in blood and mucosal epithelial tissues, and the expression level was the lowest in muscle tissue.
引文
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[12] 袁媛,宋婷,黄蓓,等.凋亡相关斑点样蛋白ASC真核表达质粒的构建及表达[J].军事医学科学院院刊,2010,34(4):328-330.
[13] YE J,YU M,ZHANG K,et al.Tissue-specific expression pattern and histological distribution of NLRP3 in Chinese yellow chicken[J].Veterinary Research Communications,2015,39(3):171-177.
[14] XIE J,BELOSEVIC M.Functional characterization of apoptosis-associated speck-like protein (ASC) of the goldfish (Carassius auratus L.)[J].Developmental & Comparative Immunology,2016,65:201-210.
[2] 孙龙华,安利国,杨桂文.凋亡相关斑点样蛋白的研究进展[J].生命科学,2012,24(4):328-333.
[3] MASUMOTO J,TANIGUCHI S,AYUKAWA K,et al.ASC,a novel 22-kDa protein,aggregates during apoptosis of human promyelocytic leukemia HL-60 cells[J].The Journal of Biological Chemistry,1999,274(48):33835-33838.
[4] 张艳.凋亡相关点样蛋白(ASC)在哮喘小鼠气道炎症中作用研究[D].广州:南方医科大学,2014.
[5] 刘文俊,邱金辉,阳佑天,等.鹅Caspase-1基因的克隆与序列分析[J].广东农业科学,2017,44(12):144-150.
[6] 刘文俊,黄植霞,朱晓琳,等.鹅GAPDH实时荧光定量PCR方法的建立[J].中国家禽,2018,40(3):10-14.
[7] 方仁东,巫芮,申艳娜.炎性小体中ASC斑点形成机理及调控机制[J].微生物学报,2016,56(9):1406-1414.
[8] SAKAIZAWA T,MATSUMURA T,FUJII C,et al.Potential role of ASC,a proapoptotic protein,for determining the cisplatin susceptibility of lung cancer cells[J].Tohoku Journal of Experimental Medicine,2018,244(2):133-144.
[9] SONG N,LI T.Regulation of NLRP3 inflammasome by phosphorylation[J].Frontiers in Immunology,2018,9:2305-2305.
[10] LI Y,HUANG Y,CAO X,et al.Functional and structural characterization of zebrafish ASC[J].FEBS Journal,2018,285(14):2691-2707.
[11] 樊钰莹,华思红,王志鹏,等.猪ASC及Ub基因的克隆及真核表达重组质粒的构建[J].动物医学进展,2019,40(1):1-5.
[12] 袁媛,宋婷,黄蓓,等.凋亡相关斑点样蛋白ASC真核表达质粒的构建及表达[J].军事医学科学院院刊,2010,34(4):328-330.
[13] YE J,YU M,ZHANG K,et al.Tissue-specific expression pattern and histological distribution of NLRP3 in Chinese yellow chicken[J].Veterinary Research Communications,2015,39(3):171-177.
[14] XIE J,BELOSEVIC M.Functional characterization of apoptosis-associated speck-like protein (ASC) of the goldfish (Carassius auratus L.)[J].Developmental & Comparative Immunology,2016,65:201-210.