GST-SRY融合蛋白原核表达和纯化

张小玉; 段君君; 王江; 贾霄; 汪小波; 何颖红

GST-SRY融合蛋白原核表达和纯化

张小玉^1,2,
段君君^1,2,
王江^1,2,
贾霄^1,2,
汪小波^1,2,
何颖红^1,2

1. 大理大学基础医学院
2. 云南省高校细胞生物学重点实验室

基金项目:

基金：国家自然科学基金资助项目 (31360288);

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出版历程
- 收稿日期: 2017-11-20

Prokaryotic Expression and Purification of GST-SRY Fusion Protein

Zhang Xiao Yu ^1,2,
Duan Jun Jun ^1,2,
Wang Jiang ^1,2,
Jia Xiao ^1,2,
Wang Xiao Bo ^1,2,
He Ying Hong ^1,2

Funds:

基金：国家自然科学基金资助项目 (31360288);

摘要

摘要: 目的构建GST-SRY融合蛋白原核表达载体, 在E.coli中诱导其表达, 并进行纯化.方法 PCR扩增SRY基因, 将SRY基因插入原核表达载体p GEX-6P-1, 经酶切和测序验证后, p GEX-6p-1-SRY转化到原核表达菌株E.coli Rosetta DE3, IPTG诱导表达GST-SRY融合蛋白, 用GST标签纯化树脂对其进行纯化.结果经双酶切和测序证实p GEX-6P-1-SRY构建正确.重组质粒转化到Rosetta DE3经IPTG诱导后表达了分子质量单位约为49 KD的重组蛋白.结论 p GEX-6P-1-SRY原核表达载体构建成功, SRY蛋白成功诱导表达并纯化, 为今后深入研究SRY蛋白的功能奠定了实验基础.
- SRY /
- 基因重组 /
- 基因表达 /
- 蛋白纯化
Abstract: Objective To construct GST-SRY fusion protein expression vector and induce its expression in E.coli, then purificate so as to obtain GST-SRY fusion protein. Me thods SRY gene was amplified by PCR and cloned into prokaryotic expression vector p GEX-6 P-1. p GEX-6 P-1-SRY plasmid was verified by restriction enzyme digestion and DNA sequencing. After that, the recombinant plasmid was transformed into E.coli Rosetta DE3.Expression of GST-SRY fusion protein was induced by IPTG, which was purified by Glutathione Sepharose later on.Re s ults Restriction enzyme digestion and sequencing demonstrated that p GEX-6 P-1-SRY was successfully obtained. The result of SDS-PAGE assay indicated that SRY was successfully expressed in E.coli Rosetta DE3 and expressed a 49 KD SRY protein.Conclus ion The successful construction of SRY gene recombinant prokaryotic expression vector and expression of SRY protein will be helpful for further studies of SRY gene function.
- SRY /
- DNA recombination /
- Gene expression /
- Protein purification

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参考文献(15)

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