肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域研究  

Interacting Domain between ABCE1-encoding Protein and the Cytoskeleton Protein Actin

作　　者：韩旭[1] 于潜[1] 田野[1] 赵希彤[1] 张磊[1] 周镝 田大力[1] HAN Xu;YU Qian;TIAN Ye;ZHAO Xitong;ZHANG Lei;ZHOU Di;TIAN Dali(Department of Thoracic Surgery,The Fourth Affiliated Hospital,China Medical University,Shenyang 110032,China)

机构地区：[1]中国医科大学附属第四医院胸外科,沈阳110032

出　　处：《中国医科大学学报》2018年第8期678-681,共4页Journal of China Medical University

基　　金：国家自然科学基金(30973502;30170914);辽宁省教育厅科学研究一般项目(L2013302)

摘　　要：目的明确肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域。方法通过转化技术,在E.coli BL21(DE3)中诱导表达前期研究获得的表达载体pGEX-4T-1-ABCE1-55,获得GST-ABCE1-55重组融合蛋白表达,应用GST pull-down的方法验证p GEX-4T-1-ABCE1-55与β-肌动蛋白有无相互作用。结果 GST-ABCE1-55是带有GST标签的含有357个氨基酸的重组蛋白,大小约66×10~3,在GST pull-down实验中,GST-ABCE1-55并不能与β-肌动蛋白特异结合。结论肺癌转移相关基因ABCE1编码蛋白与β-肌动蛋白的相互作用结构域应位于ABCE1蛋白第358位至第600位编码氨基酸中。Objective To investigate the interacting domain between ABCE1-encoding protein and the cytoskeleton protein actin. Methods Escherichia coli cells were induced with IPTG to produce the recombinant protein glutathione S-transferase（GST）-ABCE1-55 using the prokaryotic expression vector pGEX-4 T-1-ABCE1-55,which was previously acquired. A GST pull-down assay was conducted to verify the interaction between GST-ABCE1-55 and β-actin. Results The recombinant protein GST-ABCE1-55 contained a GST tag and was 66×10^3. There was no interaction between GST-ABCE1-55 and β-actin. Conclusion The interacting domain was predicted to be present between the 358 th and 600 th encoded amino acids.

关 键 词：ATP结合盒转运子E1 pGEX-4T-1原核表达载体 重组蛋白表达 蛋白质相互作用结构域 

分 类 号：R734.2[医药卫生—肿瘤]