小鼠DNAJB13与HK1的相互作用

发布时间：2018-04-11 04:13

  本文选题：DNAJB + 重组质粒　； 参考：《南方医科大学学报》2016年12期

【摘要】：目的探讨小鼠DNAJB13与HK1是否具有相互作用。方法应用双酶切连接方法构建p GEX-4T-1/Dnajb13原核表达载体,测序验证;重组质粒转化感受态细胞DH5α,用IPTG诱导融合蛋白GST-DNAJB13表达,采用SDS-PAGE考马斯亮蓝染色和Western blotting分析蛋白和鉴定;提取小鼠睾丸蛋白,采用GST pull down检测DNAJB13与HK1是否具有相互作用。结果成功构建p GEX-4T-1-Dnajb13重组质粒,测序结果与标准序列一致;转化重组质粒的大肠杆菌在37℃,IPTG浓度1 mmol/L诱导下高效表达融合蛋白;GST pull down检测结果阳性,显示DNAJB13与HK1存在相互作用。结论在小鼠睾丸中,DNAJB13与HK1存在相互作用,可能参与精子形成和精子运动。
[Abstract]:Objective to investigate the interaction between DNAJB13 and HK1 in mice.Methods the prokaryotic expression vector of p GEX-4T-1/Dnajb13 was constructed by double enzyme digestion and confirmed by sequencing. The recombinant plasmid was transformed into DH5 伪. The fusion protein GST-DNAJB13 was induced by IPTG. The protein was analyzed and identified by SDS-PAGE Coomassie brilliant blue staining and Western blotting.Mouse testicular protein was extracted and GST pull down was used to detect the interaction between DNAJB13 and HK1.Results the recombinant plasmid of p GEX-4T-1-Dnajb13 was successfully constructed, and the result of sequencing was consistent with the standard sequence, and the expression of fusion protein GST pull down in E. coli transformed by 1 mmol/L at 37 鈩，

本文编号：1734374