本文選題：DNAJB + 重組質粒��； 參考：《南方醫(yī)科大學學報》2016年12期

【摘要】：目的探討小鼠DNAJB13與HK1是否具有相互作用。方法應用雙酶切連接方法構建p GEX-4T-1/Dnajb13原核表達載體,測序驗證;重組質粒轉化感受態(tài)細胞DH5α,用IPTG誘導融合蛋白GST-DNAJB13表達,采用SDS-PAGE考馬斯亮藍染色和Western blotting分析蛋白和鑒定;提取小鼠睪丸蛋白,采用GST pull down檢測DNAJB13與HK1是否具有相互作用。結果成功構建p GEX-4T-1-Dnajb13重組質粒,測序結果與標準序列一致;轉化重組質粒的大腸桿菌在37℃,IPTG濃度1 mmol/L誘導下高效表達融合蛋白;GST pull down檢測結果陽性,顯示DNAJB13與HK1存在相互作用。結論在小鼠睪丸中,DNAJB13與HK1存在相互作用,可能參與精子形成和精子運動。
[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 偽. 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，

本文編號：1734374