摘要
目的:在大肠杆菌中表达KOX12蛋白,为进一步研究它的功能奠定基础。方法:根据GenBank中KOX12的基因序列设计相应的引物,用RT-PCR的方法从肺癌细胞株中扩增得到KOX12片段,将它克隆到PcDNA3.1F2/FOXM1质粒,然后经BamHI和XhoI双酶切,得到修饰过的KOX12基因片段,构建原核表达质粒PQE30/KOX12,以大肠杆菌M15为宿主菌表达目的蛋白,并采用Ni-NTA agarose亲和层析对目的蛋白进行了纯化。结果:构建了PQE30/KOX12原核表达体系,并检测到KOX12蛋白的有效表达。结论:成功地构建了原核表达质粒PQE30/KOX12并表达了KOX12蛋白,为进一步研究它的功能奠定了基础。
Objective To express the KOX12 protein in E.Coli for further studying its function.Methods To design the corresponding primer according to KOX12 gene sequences in the GenBank,we amplified it＇s fragments with RT-PCR from lung cancer cells.The fragments was cloned into PcDNA3.1F2/FOXM1 plasmid.The modified KOX12 gene fragment was obtained with BamHI and XhoI double digestion to construct prokaryotic expression plasmid PQE30/KOX12.The target protein was expressed into E.coli host strain M15,and was purified through Ni-NTA agarose affinity chromatography.Results We constructed the PQE30/KOX12 prokaryotic expression system,and detected effective KOX12 protein expression.Conclusion We successfully constructed prokaryotic expression plasmid PQE30/KOX12 and expressed KOX12 protein in E.coli for further study.
出处
《放射免疫学杂志》
CAS
2009年第5期524-526,共3页
Journal of Radioimmanology
