Expression of cre gene in Escherichia coli and bioassay its expression product.
- Author:
Li-Xia WANG
1
;
Zhu-Qiang ZHANG
;
Xiao-Qian HU
;
Yuan-Lei HU
;
Yin GAO
;
Zhong-Ping LIN
Author Information
1. National Key Laboratory of Protein Engineering and Plant Molecular Biology of Peking University, China.
- Publication Type:Journal Article
- MeSH:
Chromatography, DEAE-Cellulose;
Escherichia coli;
genetics;
Gene Expression Regulation, Enzymologic;
Green Fluorescent Proteins;
Integrases;
genetics;
metabolism;
Luminescent Proteins;
genetics;
metabolism;
Plasmids;
genetics;
Recombinant Proteins;
isolation & purification;
metabolism;
Viral Proteins;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2002;18(4):497-500
- CountryChina
- Language:Chinese
-
Abstract:
The Cre recombinase from bacteriophage P1 can recognize specific DNA sequences, cleave DNA at specific target sites, and then ligate it to the cleaved DNA of a second site. In this study, cre gene was cloned into the pGEM-T Easy vector via PCR procedure. Then the cre gene was inserted into an expression vector pET-29a and expressed in E. coli BL21 (DE3). A 38 kD soluble protein was expressed and named CRE. CRE was purified by DEAE-52 chromatography. Bioassay of the partially purified product showed that CRE can cleave the plasmid pGLGFP which contains two loxP site with the same direction.
