|
|
|
| Construction of Prokaryotic Expression Vector of Bovine Nanog Gene and Its Expression in E.coli |
| 广西壮族自治区南宁市大学路100号 广西大学东校园动物科技学院 邮编：530004 |
|
|
|
|
Abstract The present study is to construct prokaryotic expression vector of bovine Nanog gene and to induce its expression in E.coli (JM109). By means of reverse transcription –polymerase chain reaction(RT-PCR), cDNA of bovine Nanog gene was amplyfied from the tissues of fetal primodial genital ridges at age of six weeks. It was inserted into PMD18-T vector , then subcloned into vector pGEX-KG by gene recombination technique. After being confirmed by restriction enzyme digestion and sequencing, the recombinant plasmid pGEX-KG -Nanog was induced by IPTG（0.1-2m M/L）to express in JM109 at 25, 30, and 37℃. The results showed that（1）IPTG concentration and culture temperature exerted little impact on the expression yield of GST- Nanog fusion protein in JM109；(2) as indicated by western blotting , expressed efficiently in JM109 and reactivated with GST antibody, the fusion protein is about 60KD, which can be used to prepare polyclonal antibody of bovine Nanog gene.
|
|
Received: 23 November 2007
|
|
|
|
| [1] |
LI Zong-Shuai, SHEN Pei-Lei, LIU Lei-Lei, YANG Yang, LI Hai-Jiang, ZHANG Quan-Wei,GONG Ji-Shang, ZHAO Xing-Xu, ZHANG Yong. Obtained Transfection Cell Line for iPSCs Isolation from Bactrian Camels (Camelus bactrianus) via CRISPR/Cas9[J]. 农业生物技术学报, 2019, 27(1): 180-190. |
|
|
|
|
