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抗人端粒酶逆转录酶单链抗体基因在烟草中的表达

Expression of Anti-hTERT ScFv in Transgenic Tobacco Plants

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【作者】 董皓林李威朱向莹林娟唐克轩任大明

【Author】 DONG Hao-lin,LI Wei,ZHU Xiang-ying,LIN Juan,TANG Ke-xuan,REN Da-ming(State Key Laboratory of Genetic Engineering,Institute of Genetics,Fudan University,Shanghai 200433 China)

【机构】 复旦大学遗传学研究所遗传工程国家重点实验室复旦大学遗传学研究所遗传工程国家重点实验室 上海200433上海200433

【摘要】 构建了含抗人端粒酶逆转录酶单链抗体基因ScFv-hTERT的植物表达载体p1304-SH,通过农杆菌介导的叶盘法转化烟草.转基因烟草植株叶片总DNA的PCR,Southern b lot检测结果表明,ScFv-hTERT基因已整合进了转基因烟草植株基因组中;RT-PCR,SDS-PAGE分析证实目的基因已在烟草叶片中成功表达,竞争性ELISA的结果表明,表达的重组抗体与其抗原有良好的结合活性.

【Abstract】 Anti-hTERT single chain Fv was cloned into a modified plant expression vector pCAMBIA1304 to generate a designated p1304-SH.Then recombinant plasmid was transformed into tobacco using leaf disk mediated by Agrobacterium tumefaciens EHA105.PCR and Southern blot were used to confirm the integration of the gene in the transgenic plant genome.RT-PCR and SDS-PAGE analysis showed that the ScFv-hTERT gene was successfully expressed in the transgenic tobacco.The result of competitive ELISA indicated that the gene product has activity of binding with its antigen.

  • 【文献出处】 复旦学报(自然科学版) ,Journal of Fudan University(Natural Science) , 编辑部邮箱 ,2006年06期
  • 【分类号】Q943.2
  • 【被引频次】4
  • 【下载频次】126
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