Construction,prokaryotic expression and purification of FimH1-156 fusion protein
10.3969/j.issn.1671-8348.2014.05.029
- VernacularTitle:FimH1-156融合基因的构建及其原核蛋白表达与纯化
- Author:
Shiwei YIN
;
Liyun ZOU
;
Ying ZHANG
;
Jinyu ZHANG
;
Sha TANG
;
Weiwei SHI
;
Yuzhang WU
;
Fahuan YUAN
;
Jingbo ZHANG
- Publication Type:Journal Article
- Keywords:
FimH1 -156;
fusion protein;
expression;
purification
- From:
Chongqing Medicine
2014;(5):589-591
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and express a prokaryotic expression vector carrying the gene of FimH 1-156 that comprises human lysosome membrane protein 2 P41-49 gene ,and to express and purify the fusion protein .Methods FimH1-156 gene was cloned from plasmid pPKL241 by PCR ,and inserted into vector pET-28a(+ ) to obtain prokaryotic expression plasmid pET-28a-FimH . After transforming Escherichia coli BL21(DE3) with pET-28a-FimH ,fusion protein FimH1-156 was expressed under induction .The target fusion protein was purified ,and its antigenicity was detected through Western blot .Results The expressed recombinant pro-tein was purified ,the expression of protein was the highest when IPTG was 1 mmol/L and 4h after induction ,it was expressed as include body form ,and the expressed protein was identified to react with monoclonal antibodies 6 × His by Western blotting .Conclu-sion We cloned FimH1-156 fusion protein expressed genes successfully ,constructed prokaryotic expression vector ,and won the in-clusion body purification of FimH1-156 fusion protein .
