细胞角蛋白18真核表达载体的构建及其对NF-κB信号通路的影响  

Construction of a eukaryotic vector expressing CK18 gene and its effect on NF-κB pathway

作　　者：姚景辉[1,2,3] 王建[2,3] 王晓辉[2,3] 黄晓碧[1,2,3] 杨晓明[2,3] 汪思应[1] 

机构地区：[1]安徽医科大学病理生理学教研室,合肥230032 [2]军事医学科学院放射与辐射医学研究所 [3]北京(国家)蛋白质组研究中心,北京100850

出　　处：《安徽医科大学学报》2009年第2期150-153,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学创新群体基金资助项目(编号:30621063);国家高新技术研究发展计划(863计划)资助项目(编号:2006AA02A310);国家重点基础研究发展规划(973计划)资助项目(编号:2006CB910802);安徽省人才开发基金资助项目(编号:2002Z035)

摘　　要：目的构建人细胞角蛋白18(CK18)真核表达载体,以研究CK18与NF-κB信号通路的关系。方法经PCR扩增CK18基因片段,用pCMV-Myc及pDsRed1-N1载体构建真核重组质粒,瞬时转染HEK293细胞,用Western blot方法检测pCMV-Myc-CK18的表达,在荧光显微镜下观察pD-sRed1-N1-CK18的定位情况。用双荧光素酶报告基因系统,检测CK18对NF-κB信号通路的影响。结果构建了人CK18真核表达载体,发现CK18定位于细胞质,并抑制NF-κB的转录活性。结论构建了人CK18真核表达载体,CK18负调控NF-κB信号通路。Objective Construct the recombinant eukaryotic vectors expressing CK18cDNA, and explore the correlation between CK18 gene and NF-κB. Methods The fragment of CK18 gene was amplified by polymerase chain reaction（ PCR）, eukaryotic vectors expressing CK18 were constructed with pCMV-Myc or pDsRedl-N1, and transfected into HEK293 cells. The expression of CK18 was detected by Western blot, and observed the cells with fluorescent microscope. CK18 how to influence NF-κB was detected by luciferase system. Results Recombinant plasmids pCMV-Myc-CK18 and pDsRedl-N1-CK18 were successfully constructed and transfected into HEK293 cells. CK18 localized in the cytoplasm. CK18 inhibits activation of NF-κB. Conclusion CK18 negatively regulates NF-κB activation.

关 键 词：NF-ΚB 基因表达 

分 类 号：R329.4[医药卫生—人体解剖和组织胚胎学] R363.1[医药卫生—基础医学]