hTERT^(1658-2070)在大肠杆菌中的克隆及表达  

Cloning and expression of hTERT^(1658-2070) in E. Coli

作　　者：童茵[1] 金洁[1] 娄引军[1] 

机构地区：[1]浙江大学血液病研究所,浙江310003

出　　处：《肿瘤》2002年第4期276-278,共3页Tumor

基　　金：浙江省自然科学基金资助项目 (3 0 0 5 0 7) ;浙江省科技厅国际合作项目 (0 0 110 613 0 ) ;国家教育部留学回国人员科研启动基金资助项目

摘　　要：目的　用基因工程的方法在大肠杆菌中表达人端粒酶逆转录酶 (hTERT)部分活性片段。方法　PCR扩增hTERTcDNA16 5 8 2 0 70片段 ,将其克隆至表达载体pGEX 5x 3上 ,转化大肠杆菌TG1,获得hTERT蛋白片段表达。并经DNA测序及Westernblot验证表达蛋白。结果　表达出的蛋白为GST hTERT融合蛋白 ,分子量为 38.8kD。经DNA测序及Westernblot证实表达蛋白即为所需蛋白。结论　在大肠杆菌中克隆并表达了hTERT部分片段。Objective To experss a fragment of human telomerase reverse transcriptase (hTERT) with special function in E. Coli by genetic engineering. Methods A fragment of hTERT cDNA 1658 2070 was amplified by PCR method and was cloned into expression plasmid. Then the recombinant plasmid was transformed into E. Coli strain TG1 and the expression strain was obtained. DNA sequencing and Western blot was used to characterize the expressed protein. Results The experessed protein was GST hTERT fusion protein with the molecular mass of 38.8 kDa. It was confirmed by DNA sequencing and Western blot. Conclusion The fragment of hTERT cDNA was cloned and expressed in E.Coli.

关 键 词：人端粒酶逆转录酶 基因克隆 蛋白表达 聚合酶链反应 大肠杆菌 

分 类 号：Q78[生物学—分子生物学]