BS69细胞原核表达载体的构建和诱导表达

Construction and Inducible Expression of Prokaryotic Expression Plasmid of BS69

目的构建BS69原核表达载体,并且对其进行初步的表达。方法将BS69克隆到原核表达载体PGEX-4T·3中,之后对重组的质粒进行鉴定;再将质粒转化入E.coli BL21中,使用IPTG诱导目的蛋白。结果重组子利用限制性内切酶进行酶切、PCR鉴定和DNA序列分析得到原核表达载体PGEX-4T·3-BS69;IPTG诱导表达后,SDS-PAGE分析显示目的蛋白在E.coli BL21中高效表达,分子量约为69k D。结论成功构建了BS69原核表达载体,完成了其在E.coli BL21中的诱导表达,为进一步在体外研究BS69提供了条件。

Objective： To construct the prokaryotic expression plasmid of PGEX- 4T·3- BS69 and express BS69 in E. coli BL21. Methods： the BS69 was subcloned into PGEX- 4T·3 vector,The recombinants were finally sequenced and identified by restrictive endonuclease digestion. Then the recombinants were transferred into E. coli BL21 for expression and induced by 1 mmol / L IPTG. The expression product of BS69 gene was analyzed by SDS- PAGE. Results： PGEX- 4T·3- BS69 prokaryotic expression vectors was successfully constructed and BS69 can induced in E. coli. BL21. Molecular weight is about 69 KD. Conclusion： The BS69 prokaryotic expression vector was successfully constructed and BS69 protein has been expressed successfully. For further research in vitro BS69 provides conditions.