| 抗甲胺磷单链抗体基因的酵母表达及性质鉴定 |
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| 引用本文： | 李铁军,李德全.抗甲胺磷单链抗体基因的酵母表达及性质鉴定[J].江西农业学报,2009,21(10):1-4,9. |
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| 作者姓名： | 李铁军 李德全 |
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| 作者单位： | [1]南通职业大学,江苏南通226007 [2]南通大学,江苏南通226007 |
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| 基金项目： | 农业部农业结构调整重大技术研究专项 |
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| 摘 要： | 为了构建高效表达阳性抗甲胺磷单链抗体(scFv)基因的巴斯德毕赤酵母Pichia pastoris系统,设计引物扩增阳性克隆scFv基因,亚克隆至表达载体pPICZαC,构建重组酵母表达质粒pPICZαC-scFv,线性化pPICZαC-scFv并高效电转P.pastoris(X-33),抗性梯度筛选转化子以获得高效表达菌株,对获得的菌株先后进行表达条件的优化、优化条件下的诱导表达及单链抗体性质研究。结果:获得了稳定高效表达菌株X-33-Pp-Met-28D4-1,该菌株优化条件下的scFv诱导表达量为25 mg/L,纯化产物对甲胺磷的抑制中浓度IC50为93.52μg/mL,抗原抗体亲合常数为2.34×10-8mol/L。因此利用Pichia pastoris表达系统可大量制备功能性抗甲胺磷单链抗体。
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| 关 键 词： | 巴斯德毕赤酵母 单链抗体 表达 甲胺磷 化学免疫特性 |
Expression and Identification of Recombinant scFv Antibodies against Methamidophos in Pichia pastoris |
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| LI Tie-jun,LI De-quan.Expression and Identification of Recombinant scFv Antibodies against Methamidophos in Pichia pastoris[J].Acta Agriculturae Jiangxi,2009,21(10):1-4,9. |
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| Authors: | LI Tie-jun LI De-quan |
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| Institution: | LI Tie-jun1,LI De-quan2(1.Nantong Vocational University,Nantong 226007,China,2.College of Life Sciences,Nantong University,China) |
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| Abstract: | In order to high-effectively express a positive single-chain Fv(scFv) gene against methamidophos in Pichia pastoris,the special scFv gene was amplified from a positive clone and then sub-cloned into the expression vector pPICZa C;the constructed recombinant plasmid pPICZa C-scFv was linearized and transformed into P.pastoris(X-33);the transformant which showed strong expression of antibodies was isolated,and its culture conditions were optimized;the immunochemical characteristics of the scFv antibodies prod... |
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| Keywords: | Pichia pastoris scFv Expression Methamidophos Immunochemical characteristics |
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