Memo蛋白的融合表达和纯化  

Fusion Expression and Purification of Memo Protein

作　　者：宁康[1] 张浩[1] 丁丽华[1] 王晓辉[1] 范洪学[2] 李杰之[1] 李熔[1] 叶棋浓[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]吉林大学公共卫生学院,吉林长春130021

出　　处：《生物技术通讯》2006年第4期560-562,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(30530320;30500091;30428012)

摘　　要：目的:一些临床预后较差的恶性肿瘤一般具有很高的转移性和侵袭性,近来的研究表明,Memo蛋白与恶性肿瘤的这些特性有十分密切的关系。本研究拟克隆、融合表达和纯化Memo蛋白。方法:用PCR方法从乳腺文库中扩增Memo蛋白编码序列,并将其以正确相位与pGST载体中的GST编码序列融合,构建成重组质粒pGST-Memo。将此重组质粒转化大肠杆菌DH5α后,用谷胱甘肽-Sepharose4B纯化融合蛋白,并用Western印迹检测融合蛋白的表达。结果:构建了Memo蛋白的融合表达载体,并在大肠杆菌DH5α中得到表达,经谷胱甘肽-Sepharose4B亲和层析获得了纯化的GST-Memo融合蛋白。Western印迹检测表明,此蛋白可与GST抗体反应。结论:Memo蛋白的克隆、融合表达和纯化,为进一步研究恶性肿瘤的转移性、侵袭性及其相关机理提供了有用的材料。Objective： Clinically, some malignant tumors with poor prognosis are characteristic of high metastasis and invasiveness. Recent studies showed that Memo protein associates with the outcome of cancer patients. To constructed pGSTMemo recombinant plasmid and to purify the fusion protein of GST-Memo. Methods： The coding sequence of Memo protein was amplified by PCR and fused in frame with the coding region of GST in the pGST vector to generate the pGSTMemo recombinant plasmid. The fusion protein GST-Memo was expressed in E.coli DH5α and purified by GlutathioneSepharose 4B affinity chromatography. The fusion protein was characterized by Western blot. Results： The pGST-Memo recombinant plasmid was successfully obtained and the fusion protein was purified. Western blot analysis showed that the fusion protein interacts with GST antibody. Conclusion： This study provides useful materials for investigation into Memoassociated mechanisms of cancer metastasis and invasiveness.

关 键 词：Memo蛋白 克隆 融合表达 

分 类 号：Q786[生物学—分子生物学]