本文選題：DNAJB + 重組質(zhì)粒　； 參考：《南方醫(yī)科大學(xué)學(xué)報(bào)》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(yīng)用雙酶切連接方法構(gòu)建p GEX-4T-1/Dnajb13原核表達(dá)載體,測序驗(yàn)證;重組質(zhì)粒轉(zhuǎn)化感受態(tài)細(xì)胞DH5α,用IPTG誘導(dǎo)融合蛋白GST-DNAJB13表達(dá),采用SDS-PAGE考馬斯亮藍(lán)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(jié)果成功構(gòu)建p GEX-4T-1-Dnajb13重組質(zhì)粒,測序結(jié)果與標(biāo)準(zhǔn)序列一致;轉(zhuǎn)化重組質(zhì)粒的大腸桿菌在37℃,IPTG濃度1 mmol/L誘導(dǎo)下高效表達(dá)融合蛋白;GST pull down檢測結(jié)果陽性,顯示DNAJB13與HK1存在相互作用。結(jié)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(yùn)動(dòng)。
[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