Survivin基因融合蛋白的表达与纯化

doi:10.3969/j.issn.1674-490X.2014.01.003

摘要/Abstract

摘要： 目的： Survivin是凋亡抑制蛋白(IAP)中最强的凋亡抑制因子，纯化得到其功能结构域(BIR)的融合蛋白可为进一步研究Survivin基因与相关基因的相互作用提供依据。方法利用PCR技术扩增Survivin基因中的BIR序列后连接到pGEX4T-2载体上，经大肠杆菌(DH5α)验证后，转化BL21大肠杆菌菌株并用IPTG进行诱导，以谷胱甘肽琼脂糖亲和层析技术纯化融合蛋白。结果实验扩增得到的BIR片段约为276 bp，纯化得到的融合蛋白分子质量大小约为37 ku。结论 PCR结果与预期的片段大小一致，构建重组体后经IPTG在22℃下诱导2 h，菌体中融合蛋白的表达量显著增加，为下一步研究Survivin基因的功能和与其相互作用的蛋白奠定了基础。

关键词: Survivin, pGEX4T-2, IPTG诱导, 纯化

Abstract: Objective Survivin gene is the strongest apoptosis-inhibition factor among inhibitor of apoptosis protein (IAP), and purification of fusion protein of functional domains (baculovirus IAP repeat, BIR) may provide the foundation for interaction research between the gene and related other genes. Methods BIR sequence, after ampliifcation with PCR and validation by Escherichia coli DH5αthrough pGEX4T-2 vector, was transformed into E. coli BL21. Fusion protein of BIR was induced by IPTG and puriifed by Glutathione Agarose Affinity Chromatography. Results The amplified fragment BIR was about 276bp, purified fusion protein molecular weight of approximately 37ku. Conclusion PCR results are consistent with the expected fragment size, recombinant plasmids after 2h after IPTG induction, the cell in the fusion protein increase signiifcantly, which provide basis for further research on Survivin gene function and its interacting protein.

Key words: Survivin, pGEX4T-2, IPTG induction, puriifcation

中图分类号:

李丽,刘晶,蒋继志. Survivin基因融合蛋白的表达与纯化[J]. 医学研究与教育, 2014, 31(1): 13-16.

LI Li,LIU Jing,JIANG Jizhi. Expression and puriifcation of Survivin gene fusion protein[J]. Medical Reserch and Education, 2014, 31(1): 13-16.

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