色素上皮细胞衍生因子原核表达重组体的构建  

Construction of recombination of E. coli DH5 of human pigment epithelium derived factor

作　　者：杨楠[1] 马颖哲[1] 张桂荣[1] 杨翰仪[1] 

机构地区：[1]吉林大学基础医学院生物化学教研室,吉林长春130021

出　　处：《中国老年学杂志》2005年第5期512-514,共3页Chinese Journal of Gerontology

摘　　要：目的构建色素上皮细胞衍生因子(PEDF)原核表达重组体。方法从胚胎绒毛中提取总RNA,经RTPCR扩增,获得编码人PEDF的全基因序列,采用TA克隆法,将PEDF基因克隆入pMD18T载体中并转化入E.coliJM109,经DNA测序正确后构建原核表达重组体pET28aPEDF,转化E.coliDH5a。结果经酶切及DNA测序鉴定,获得了PEDF原核表达重组体pET28aPEDF。结论构建了PEDF原核表达重组体pET28aPEDF,为获得重组PEDF奠定了基础。Objective To construct recombination of E. coli DH5 of human recombinant pigment epithelium derived factor (PEDF). Methods Total RNA extracted from human embryo was amplified by RT-PCR to obtain complete gene sequences encoding human PEDF. PEDF gene was cloned into vector PMD18-T by T-A methods and transformed into E. coli JM109 to construct recombinant pMD18-PEDF after DNA sequencing and then transformed E. coli DH5α. Results The recombinant plasmid pET28a-PEDF was obtained by digestion and sequencing. Conclusions The recombinant plasmid pET28a-PEDF constructed could lay the foundation for recombinant PEDF.

关 键 词：色素上皮细胞衍生因子 克隆 载体 

分 类 号：Q784[生物学—分子生物学]