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目的:制备BCR/ABL OD结构域-HIS的重组蛋 白.方法:用RT-PCR方法扩增BCR/ABL OD结构域的基因 片段,测序正确后将其克隆入原核表达载体pET16b中,并进 行酶切鉴定,鉴定正确后转化大肠杆菌BL21,构建表达His- OD融合蛋白的菌株.经IPTG诱导表达后,镍 次氮基三乙酸 (Ni NTA)琼脂糖进行亲和层析纯化,用SDS-PAGE对该融 合蛋白的表达产率及蛋白纯度进行了分析.结果:获得了 His-OD重组融合蛋白,纯度在95%以上,产物得率约75%. 结论:成功制备高纯度His-OD融合蛋白,为进一步研究OD 结构域的生物学功能奠定了基础.
OBJECTIVE: To prepare the recombinant protein of BCR / ABL OD domain-HIS.Methods: The gene fragment of BCR / ABL OD domain was amplified by RT-PCR and cloned into the prokaryotic expression vector pET16b after sequencing, After identification and identification, the recombinant plasmid was transformed into E.coli BL21 to construct a strain expressing His-OD fusion protein.After induced by IPTG, Ni-NTA agarose was purified by affinity chromatography and analyzed by SDS-PAGE The recombinant protein His-OD was purified with the purity above 95% and the yield of the product about 75% .Conclusion: The high-purity His-OD fusion protein , Which laid the foundation for further study on the biological function of OD domain.