摘要
以人附睾cDNA为模板,PCR扩增RNase10基因,构建pET32b(+)-RNase10体外表达载体,转化大肠杆菌(Escherichia coli)感受态细胞DH5α,挑选阳性克隆,提取质粒转化BL21(DE3),IPTG诱导RNase10基因的表达,采用SDS-PAGE法检测表达产物,结果显示融合蛋白分子量为24 ku,与预期相符,表明人附睾RNase10基因在大肠杆菌中成功表达。
Using human epididymis cDNA as template,RNase10 gene was amplified and recombined into prokaryotic expression vector pET32b(+),which was constructed and transformed into Escherichia coli DH5α.Plasmids were extracted from positive clones,and transformed into BL21(DE3).Furthermore,the protein expression of RNase10 was induced by IPTG.A band about 24 ku was detected by SDS-PAGE,showing that RNase10 of human epididymis was expressed successfully in cell of E.coli DH5α.
出处
《湖北农业科学》
北大核心
2013年第17期4245-4247,共3页
Hubei Agricultural Sciences
基金
烟台大学青年基金项目(SM11Z7)
