| 抗人肝癌MDscFV基因的表达及其免疫活性初步研究 |
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| 引用本文： | 陆东东,张锡然. 抗人肝癌MDscFV基因的表达及其免疫活性初步研究[J]. 临床肝胆病杂志, 2003, 19(5): 302-303 |
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| 作者姓名： | 陆东东 张锡然 |
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| 作者单位： | 1. 南京师范大学生命科学院,江苏,南京,210097;启东肝癌研究所,江苏,启东,226200
2. 南京师范大学生命科学院,江苏,南京,210097 |
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| 摘 要： | 将抗人肝癌单克隆抗体MD的重链可变区VH和轻链可变区VL基因重组，并使其表达。采用PT-PCR技术扩增VH及VL基因，通过重叠延伸拼接PCR在VH和VL基因间引入连接短肽，体外构建MDscFV基因，经过常规转化与筛选，诱导表达蛋白。MDscFV基因全长为732bp，VH354bp位于上游VL330bp位于下游。重组蛋白相对分子量36kDa。scFV保留了与亲本抗体MD相似的免疫活性。成功地构建了抗人单链抗体MDscFV，并获得了较高水平的功能性表达。
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| 关 键 词： | 人肝癌 MDscFV基因 基因表达 免疫活性 基因重组 单克隆抗体MD |
| 文章编号： | 1001-5256(2003)05-0302-02 |
| 修稿时间： | 2002-09-24 |
Expression of single Chain Fv gene against human hepatocellular carcinoma and its immunoreactivity |
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| LU Dong -dong',Zhang Xi - ran.. Expression of single Chain Fv gene against human hepatocellular carcinoma and its immunoreactivity[J]. Chinese Journal of Clinical Hepatology, 2003, 19(5): 302-303 |
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| Authors: | LU Dong -dong' Zhang Xi - ran. |
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| Affiliation: | LU Dong -dong1'2,Zhang Xi - ran1. |
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| Abstract: | This study was designed to recombine the genes from the variable regions of light chain and heavy chain of MD, a monoclonal antibody against human liver cancer VH and VL gene were amplified, by RT- PCR, and then were connected to each other by a linker peptide using extension overlap splicing PCR to obtain the MDscFv gene. Sequence analysis showed that scFv gene consisted of 831bp, among them, 404bp for heavy chain gene, located upstream of scFv gene, and 400bp for the light chain gene, located downstream. The relative molecular weight of fusion protein is 36kDa which was consistent with the theoretically predicted value. ELISA analysis revealed that scFv had equal immunoreactivity to the parent MD antibody. MDscFv gene against human hepatocellular carcinoma was successfully constructed, and functionally expressed in E. coli. |
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| Keywords: | Single chain Fv(scFv) Hepatocellular carcinoma Gene expression |
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