[Expression of cre gene in Escherichia coli and bioassay its expression product].
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.
Wang
Li-Xia
LX
National Key Laboratory of Protein Engineering and Plant Molecular Biology of Peking University, China.
Zhang
Zhu-Qiang
ZQ
Hu
Xiao-Qian
XQ
Hu
Yuan-Lei
YL
Gao
Yin
Y
Lin
Zhong-Ping
ZP
D002848:Chromatography, DEAE-Cellulose; D004926:Escherichia coli / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D049452:Green Fluorescent Proteins; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D008164:Luminescent Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases