组蛋白甲基转移酶SET7的原核表达及纯化鉴定

摘要

Objective To purify and prokaryotically express the histone methyltransferase SET7.Methods The coding sequence of full length SET7 was amplified from breast library by PCR and cloned into the pGEX-KG vector.The correct recombinant plasmid was introduced into E.coli.The expressed protein was identified by SDS-PAGE.Results DNA sequencing indicated the SET7 was constructed.GST-SET7 fusion protein was identified by SDS-PAGE.Conclusion The prokaryotically expressed protein of GST-SET7 is obtained,which will falilitate further study of SET7 function.%目的：原核表达并纯化组蛋白甲基转移酶 SET7。方法以乳腺文库为模板，PCR 扩增 SET7基因，克隆到 pGEX-KG 载体，构建 pGEX-KG-SET7；经酶切鉴定后转化进行小量诱导，通过 SDS-PAGE 检测融合蛋白 GST-SET7的纯化效果。结果DNA 测序结果表明，pGEX-KG-SET7原核表达载体构建成功。SDS-PAGE 检测显示获得相对分子质量为72×103的融合蛋白。结论纯化得到原核表达的 GST-SET7融合蛋白，为进一步研究 SET7在乳腺癌中的功能奠定了基础。