Cloning and expression of rat heme oxygenase 1 in E.coli / 吉林大学学报(医学版)

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.

Cloning,prokaryotic expression and identification of human cell division cycle gene 2 / 吉林大学学报(医学版)

Objective To clone human cell division cycle gene 2(CDC2)from human liver cancer tissue and express CDC2 protein in E.coli BL-21(DE3).Methods The total RNA was extracted from liver cancer tissue and amplified by reverse transcription polymerase chain reaction(RT-PCR).The PCR products were cloned into the prokaryotic expression vector pET28a(+)followed by DNA sequencing.The recombinant pET28a(+)/CDC2(rCDC2)plasmid was transformed into E.coli.The rCDC2 was induced with IPTG and characterized by SDS-PAGE.Results The cloned CDC2 gene was composed of 894 nucleotides,and is accordance with that reported in GenBank.The prokaryotic expression vector was constructed successfully.The CDC2 protein was successfully expressed in E.coli.Conclusion The CDC2 cDNA is successfully cloned from human liver cancer tissue,and can express in E.coli.