Construction and identification of recombinant vector encoding hTERT and HSP70 genes
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    Abstract:

    Objective:To construct the fusion gene of human hTERT and human heat shock protein 70(HSP70), and expressed it in E.coli. Methods:The hTERT and HSP70 genes were amplified by PCR. The hTERT gene was digested by NcoⅠand EcoRⅠ enzyme and inserted into the plasmid pET32a to construct pET32a-hTERT plasmid. Then the HSP70 gene was cloned into the expression vector pET32a-hTERT between EcoRⅠ and HindⅢ site to construct the prokaryotic expression plasmid pET32a-hTERT-HSP70. The E.coli BL21 contained the plasmid was induced by IPTG. Results:The HSP70 gene and hTERT gene segments were amplified from their templates; the DNA sequencing result showed that the prokaryotic fusion expression plasmid pET32a-hTERT-HSP70 was successfully constructed. The E.coli BL21 contained the plasmid could express a Mr 110 kD protein after induced by IPTG. Conclusion:The successful expression of fusion protein of human hTERT and heat shock protein 70 enables the further research of the HSP70 protein as a adjuvant-free carrier.

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唐鹿群,张 蕾,郭人花,刘 丰,苏 川,束永前.人端粒酶逆转录酶与热休克蛋白70融合表达载体的构建及原核表达[J].南京医科大学学报(自然科学版英文版),2007,(8):825-828.

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  • Received:December 09,2006
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