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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GSTSET7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。
Objective To express and purify histone methyltransferase SET7 in prokaryotic cells. Methods SET7 gene was amplified by PCR from the mammary gland. The recombinant plasmid pGEX-KG-SET7 was cloned into pGEX-KG vector. The recombinant plasmid pGEX-KG-SET7 was transformed into E.coli BL21. Results DNA sequencing results showed that pGEX-KG-SET7 prokaryotic expression vector was successfully constructed. SDS-PAGE showed that the relative molecular mass of 72 × 103 fusion protein. Conclusion The prokaryotic expression of GST-SET7 fusion protein was purified, which laid the foundation for the further study on the function of SET7 in breast cancer.