Cloning and expression of rat heme oxygenase 1 in E.coli
- VernacularTitle:大鼠血红素氧化酶1的基因克隆和原核表达
- Author:
Bo XU
;
Yongchen ZHENG
;
Shaoyang FEI
;
Hang GAO
- Publication Type:Journal Article
- Keywords:
heme oxygenase(decyclizing);
cloning,molecular;
prokaryotic expression
- From:
Journal of Jilin University(Medicine Edition)
2006;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone the rat heme oxygenase-1(RHO-1) from rat spleen and express RHO-1 in E.coli BL-21.Methods The total RNA was extracted from rat spleen and amplified by reverse transcription polymerase chain reaction(RT-PCR).PCR products were cloned into pMD18-T(TA)vector followed by DNA sequencing.RHO-1 cDNA fragments in TA vector were subcloned into the prokaryotic expression vector pET28a(+).The recombinant pET28a(+)/RHO-1(rRHO-1) plasmid was transformed into E.coli.The rRHO-1 was induced with IPTG and characterized by SDS-PAGE.Results The cloned RHO-1 gene was composed of 870 nucleotides,and was accordance with the sequence reported in GenBank.The prokaryotic expression vector was constructed successfully.The RHO-1 protein was successfully expressed in E.coli.Conclusion The prokaryotic expression vector of rRHO-1 has been constructed,and the fusion protein has been successfully expressed.
