Increasing Expression of an Anti-Picloram Single-Chain Variable Fragment (ScFv) Antibody and Resistance to Picloram in Transgenic Tobacco (Nicotiana tabacum)

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• 作者：Fernando Olea-Popelka ; Michael D. McLean ; Jonathan Horsman ; Kurt Almquist ; Jim E. Brandle ; and J. Christopher Hall
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2005
• 出版时间：August 24, 2005
• 年：2005
• 卷：53
• 期：17
• 页码：6683 - 6690
• 全文大小：545K
• 年卷期：v.53,no.17(August 24, 2005)
• ISSN：1520-5118

文摘

Systematic research involving four chimeric gene constructions designed to express the same anti-picloram single-chain variable fragment (scFv) antibody is described. Agrobacterium-mediatedtransformation produced at least 25 transgenic tobacco plants with each of these, and the number ofT-DNA loci in each plant was determined using kanamycin-resistance segregation assays. The relativeamounts of active and total scFv in each plant were evaluated using quantitative enzyme-linkedimmunosorbent assay and immunoblot technologies, respectively. No significant differences in scFvactivity were found among the four groups of single-locus plants, although the 35S/M construct wasfound to produce significantly more total anti-picloram scFv than the other three constructs. A dose-response bioassay involving T₁ seedlings from several of the highest expressers of active scFvdemonstrated resistance to a constant exposure of picloram at 5 × 10^-8 M. Other approaches forincreasing antibody-based herbicide resistance are discussed, as further improvements are neededbefore practical application of this technology.Keywords: Single-chain variable fragment antibody; scFv; picloram; herbicide resistance; immunomodulation; transgenic plant; tobacco; Nicotiana tabacum