分泌型肿瘤坏死因子相关激活诱导因子融合蛋白在大肠杆菌中的表达和纯化

Expression and Purification of Secreted form TNF-related Activation-induced Cytokine in E.coli as a Maltose Binding Protein Fusion

目的 表达及纯化人分泌型肿瘤坏死因子相关激活诱导因子 (secretedformTNF relatedactivation in ducedcytokine,sTRANCE)蛋白。方法 将sTRANCE结构基因重组到含麦芽糖结合蛋白 (MBP)的融合蛋白原核表达载体 pMAL c2x中 ,在大肠杆菌中进行表达。 结果 经IPTG诱导表达出的MBP sTRANCE融合蛋白相对分子质量约 70 0 0 0 ,并经Westernblot分析证实。用直链淀粉琼脂糖凝胶 (amyloseresin)亲和层析纯化得到电泳均一的融合蛋白。结论 成功地获得sTRANCE蛋白 ,为进一步研究其生物学性能和用于肿瘤生物治疗的可能性打下了基础。

Purpose To express and purify the protein of sTRANCE(secreted form TNF related activation induced cytokine). Methods sTRANCE gene was cloned into expressed vector PMAL c2x and the fusion protein MBP sTRANCE was expressed in E.coli . Results The molecular weights of MBP sTRANCE was 70 000.Western blot analysis showed the fusion protein presented the specific MBP antigenicity.MBP sTANCE fusion protein were purified by amylose resin affinity chromatography and showed to be homogeneity in SDS PAGE. Conclusions The MBP sTRANCE fusion protein was obtained and maybe suitable for studying biological functions and potential utility in neoplasm treatment.