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目的:构建含人端粒酶RNA(hTR)核酶基因的真核表达质粒pBBS212Rz,为以端粒酶为靶点的基因治疗消化系肿瘤研究奠定基础.方法:将人工合成的端粒酶核酶基因通过基因重组定向克隆插入到真核表达质粒载体pBBS212中.根据260nm的紫外光吸收值计算重组质粒pBBS212Rz浓度.结果:端粒酶核酶基因成功地定向插入了载体pBBS212,经酶切后用聚丙烯凝胶电泳鉴定确认.重组质粒浓度为1.77g/L.结论:成功构建了含人端粒酶RNA(hTR)核酶基因的真核表达重组质粒pBBS212Rz.
OBJECTIVE: To construct the eukaryotic expression plasmid pBBS212Rz containing human telomerase RNA (hTR) ribozyme gene and lay the foundation for the study of gene therapy of gastrointestinal tumors with telomerase targeting.Methods: The human telomerase nuclear The recombinant plasmid pBBS212Rz was inserted into the eukaryotic expression vector pBBS212 by gene recombination and cloned into pBBS212.Results: The plasmid pBBS212Rz was successfully inserted into vector pBBS212 by UV absorbance at 260nm, It was confirmed by the polyacrylamide gel electrophoresis that the recombinant plasmid concentration was 1.77g / L. Conclusion: The eukaryotic expression recombinant plasmid pBBS212Rz containing human telomerase RNA (hTR) ribozyme gene was successfully constructed.