Cloning and prokaryotic expression of LEN-5 β-lactamase gene
10.3969/j.issn.1002-2694.2010.03.019
- VernacularTitle:LEN-5型β-内酰胺酶基因的克隆及原核表达
- Author:
Yuemei LU
;
Ruanzhang ZHANG
;
Yuhua HU
;
Yunhua ZHONG
;
Xuecheng WU
;
Shengwen CHEN
;
Shayan WANG
- Publication Type:Journal Article
- Keywords:
β-lactamase;
prokaryotic expression;
isoelectric focusing electrophoresis
- From:
Chinese Journal of Zoonoses
2010;(3):266-268
- CountryChina
- Language:Chinese
-
Abstract:
In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.
