医学信息Issue(20)：215-216,2.
α-烯醇化酶重组质粒的构建及其表达形式的研究
Construction and Expression ofα-enol Form of the Enzyme in Clinical Study of Recombinant Plasmi
摘要
Abstract
Objective To construct an alpha enolase (ENO1) recombinant expression plasmid, and study its expression form. Methods The cDNA was synthesized using total RNA of Human Umbilical Vein Endothelial Cel (HUVEC) as the template. ENO1 gene was amplified from cDNA using specific forward and reverse primers by polymerase chain reaction (PCR). The recombinant expression vector was obtained by ligating ENO1 gene and pET28 (a) using T4 ligase. ENO1 protein was produced by transfecting expression vector pET28 (a)-ENO1 into BL21 (DE3) E.Coli. and subsequent IPTG induction. The supernatant and bacteria precipitates were collected and separated by SDS-PAGE electrophoresis, respectively, to determine the expression form of the recombinant plasmid. Results The sequence of cloned gene of ENO1 in the recombinant expression plasmid pET28 (a)-ENO1 was consistent with the sequence from the GeneBank; the expression form of PET28 (a) -ENO1 is soluble expression. Conclusion A kind of recombinant expression vector, pET28 (a) - ENO1, for expressing ENO1 protein was successful y constructed, and its expression form in E.Coli was also confirmed. The studies here wil lay the foundation for further research on biological activity and function of ENO1 .关键词
图书馆Key words
Alpha enolase/Prokaryotic expression/Recombinant plasmid/Expression form引用本文复制引用
宁兴旺,谢小兵,朱惠斌,王述湘,葛金文..α-烯醇化酶重组质粒的构建及其表达形式的研究[J].医学信息,2014,(20):215-216,2.基金项目
湖南省科技厅国际合作课题，项目编号2012WK3059。 （）
