大鼠寡霉素敏感相关蛋白原核表达载体的构建

Construction and sequence analyzing of rat oligomycin sensitivity-conferring protein prokaryotic expression vector

目的:克隆大鼠寡霉素敏感相关蛋白(OSCP)基因全长cDNA,构建原核表达载体pQE30-Xa-OSCP。方法:利用逆转录-聚合酶链式反应(RT-PCR)技术从大鼠脑组织中扩增出目的基因,先以A-T连接方式克隆到pTA2载体中,再将其克隆到原核表达载体pQE30-Xa中,并进行酶切、测序鉴定。结果:RT-PCR扩增出约700bp特异性片段,重组原核表达载体经酶切鉴定结果同预期相符,经测序鉴定序列同源性与GeneBank报道一致。结论:成功构建了重组原核表达载体pQE30-Xa-OSCP。

Objective： To clone rat oligomycin sensitivity-conferring protein cDNA and to construct prokaryotic expression vector pQE30-Xa-OSCP. Methods：RT-PCR was used to amplify the target gene from the tissues of rat brain.The target gene was cloned into pTA2 and then subcloned into prokaryotic expression vector pQE30-Xa and confirmed by restriction analysis and DNA sequencing. Results：About 700 bp DNA fragment was amplified with RT-PCR. Restriction analyses of the recombinant prokaryotic expression vector was consistent with the theoretically predicted results and sequence analysis revealed the same homology to published data in GeneBank. Conclusion：The recombinant prokaryotic expression vector pQE30-Xa-OSCP is constructed successfully.