瘦蛋白基因的克隆及在大肠杆菌中的表达  

Cloning of Porcine Leptin Gene and Its Expression in E.coli

作　　者：张忠芳[1] 高士争[2] 张曦[2] 李士泽[3] 张玉静[4] 

机构地区：[1]北华大学医学院,吉林吉林132001 [2]云南农业大学云南省动物营养与饲料重点实验室,云南昆明650201 [3]黑龙江八一农垦大学,黑龙江大庆163319 [4]解放军军需大学军事兽医系,吉林长春130062

出　　处：《中国兽医学报》2004年第5期463-466,共4页Chinese Journal of Veterinary Science

基　　金：云南省自然科学基金资助项目 (2 0 0 2 C0 0 45 M)

摘　　要：采用 RT- PCR方法从猪脂肪组织扩增出瘦蛋白基因 ,将其插入 p MD1 8- T克隆载体 ,经序列分析证明 ,所获得的目的基因为 4 4 1 bp,与预期大小一致。提取质粒后用 Eco R 和 Xho 双酶切 ,克隆入 p ET- 2 8a表达载体 ,将阳性重组质粒转化表达受体菌 E.coli BL 2 1 ( DE3) ,经 IPTG诱导 ,SDS- PAGE检测 ,证明在 E.coli BL 2The porcine leptin gene was amplified by RT-PCR and the PCR product was cloned into a clone vector pMD18-T. Sequence analysis showed that the gene obtained was 441 bp long, with 99% homology with porcine leptin gene. Then the gene was subcloned into an expression vector pET-28a in correct reading-frame. The positive recombinant plasmids were then transformed into host strain E.coli BL21(DE3) and induced by IPTG. The recombinant protein was expressed in E.coli successfully as confirmed by SDS-PAGE.

关 键 词：瘦蛋白 基因 克隆 大肠杆菌 表达 脂肪细胞 

分 类 号：Q78[生物学—分子生物学] Q51