端粒酶核酶真核表达质粒pBBS212Rz的构建与鉴定

Construction and assessment of eukaryotic expression plasmid pBBS212Rz containing ribozyme gene against hTR

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.

AIM:To construct the eukaryotic expression plasmidpBBS212Rz containing ribozyme gene against humantelomerase RNA (hTR) for future study of digestive tumorgene therapy.METHODS:The synthesized ribozyme gene against hTR wasinserted into the eukaryotic expression vector pBBS212 withdefinite direction. The recombinated plasmid was calledpBBS212Rz. Its density was determined by measuring theabsorbance at 260 nm.RESULTS:Ribozyme gene was successfully inserted into theeukaryotic expression vector pBBS212. The recombinant wasproved to be the same as designed by restriction endonu-clease analysis and polyacrylamide gel electrophoresis(PAGE). Its mass concentration was 1.77 g/L.CONCLUSION:Eukaryotic expression plasmid recombinantpBBS212Rz containing ribozyme gene against hTR is suc-cessfully constructed.