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肿瘤血管导向肽与人突变IL-2融合蛋白的克隆、表达及生物学活性分析
引用本文:易颖,颜真,张英起,贺海平. 肿瘤血管导向肽与人突变IL-2融合蛋白的克隆、表达及生物学活性分析[J]. 广西医学, 2006, 28(4): 471-475
作者姓名:易颖  颜真  张英起  贺海平
作者单位:广西医科大学附属肿瘤医院基础实验部,南宁,530021;第四军医大学药学系生物技术中心,西安,710032
摘    要:目的获得具有生物学活性的与肿瘤血管导向肽(NGR)融合表达的突变人IL-2蛋白.方法将突变人IL-2与NGR的融合基因重组到表达载体pET-22b(+)并转化E.coli BL21(DE3),测序正确后,IPTG诱导表达,并对产物进行纯化、复性和鉴定.结果目的蛋白以包涵体形式存在,占菌体总蛋白的40%以上,Western blot分析显示纯化后的目的蛋白具有免疫结合活性,CTLL-2细胞增殖实验证明具有生物学活性.结论具有生物学活性的与肿瘤血管导向肽融合表达的重组人IL-2蛋白可成功地克隆并表达,为进一步的肿瘤导向治疗打下基础.

关 键 词:人突变IL-2  导向多肽  蛋白融合
收稿时间:2006-04-06
修稿时间:2006-04-06

Cloning, expression and analysis of biological activity of the fusion protein of recombinant mutant human IL-2 and a tumor-targeting peptide
YI Ying,YAN Zhen,ZHANG Ying-qi,HE Hai-ping. Cloning, expression and analysis of biological activity of the fusion protein of recombinant mutant human IL-2 and a tumor-targeting peptide[J]. Guangxi Medical Journal, 2006, 28(4): 471-475
Authors:YI Ying  YAN Zhen  ZHANG Ying-qi  HE Hai-ping
Affiliation:Experimental Department of Affiliated Cancer Hospital, Guangxi Medical University, Guangxi Nanning 530021,China;Experimental Department of Affiliated Cancer Hospital, Guangxi Medical University, Guangxi Nanning 530021,China;Experimental Department of Affiliated Cancer Hospital, Guangxi Medical University, Guangxi Nanning 530021,China;Experimental Department of Affiliated Cancer Hospital, Guangxi Medical University, Guangxi Nanning 530021,China
Abstract:Objective To obtain the fusion protein of recombinant mutant human IL-2 and a tumor-targeting peptide and study its biological activity.Methods Constructed a fusion gene of rmhIL-2 and a tumor-targeting peptide NGR cDNA and cloned into expression vector pET-22b(+). After identification by sequencing, the recombinant vector was transformed into E.coli BL21(DE3)for expression under induction of IPTG. The product of expression was purified, naturalized and identified.Results The product of expression presented in a form of inclusion body and more than 40% of total bacterial protein was the recombinant protein. The purified fusion protein rmhIL-2-NGR showed an immunogenicity on Western blot and a biological activity on CTLL-2 proliferative assay.Conclusion The fusion gene of rmhIL-2 and a tumor-targeting peptide can be successfully cloned and expressed. The expressed product shows biological activity and provids a chance for tumor targeting therapy.
Keywords:Mutant human interleukin-2   Tumor-targeting peptide   Fusion protein
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