原核表达重组质粒pUC57－CTP－PTEN的构建及鉴定

目的：合成基因CTP及PTEN基因，构建其原核表达重组质粒pUC57－CTP－PTEN，为后续研究其功能做准备。方法：合成基因CTP－PTEN，将该基因定向克隆到原核表达载体pUC57中，构建pUC57－CTP－PTEN重组质粒，转化大肠杆菌DH5α菌株，抽提质粒进行双酶切、测序鉴定后，用Western blotting检验表达。结果：测序鉴定CTP－PTEN基因合成成功。经双酶切、测序鉴定证实重组质粒pUC57－CTP－PTEN 成功转入DH5α，Western blotting检验pUC57－CTP－PTEN成功表达。结论：成功构建了重组质粒pUC57－CTP－PTEN，并在大肠杆菌DH5α中成功表达。

Construction and identification of the prokaryotic expression recombinant plasmid pUC57-CTP-PTEN

Objective:To construct prokaryotic expression recombinant plasmid pUC57 -CTP -PTEN of CTP gene and PTEN gene,and prepare for the follow-up study of its function.Methods:Gene CTP-PTEN was synthe-sized and cloned into the prokaryotic expression vector pUC57 to construct the recombinant plasmid pUC57 -CTP-PTEN.Then it was transformed into E.coli DH5α.The recombinant plasmid was identified by restriction enzyme di-gestion and DNA sequencing.The expression product was identified by Western blotting assay.Results:Gene CTP-PTEN was successfully synthesized.The recombinant plasmid pUC57 -CTP-PTEN was identified that it was suc-cessfully transformed into E.coli DH5αand expressed by restriction enzyme digestion,DNA sequencing and Western blotting.Conclusion:The recombinant plasmid pUC57 -CTP-PTEN was successfully constructed and expressed in E.coli DH5α.