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《Science & Technology Review》 2015-01
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Detecting expression of spindle protein gene INMAP in hepatoma cells by polyclonal antibody

ZHU Yan;KANG Jingyan;WANG Yu;SUN Le;ZHANG Haijiang;LIANG Qianjin;Beijing Key Laboratory of Gene Resource and Molecular Development,School of Life Sciences,Beijing Normal University;Key Laboratory for Cell Proliferation and Regulation Biology,Ministry of Education;School of Life Sciences,Beijing Normal University;No.4 High School of Lanzhou City;AbMax Biotechnology Co.,Ltd.;Beijing Health Guard Biotechnology Inc.;  
To investigate the function of INMAP of spindle protein and its role in malignant cell proliferation,an INMAP polyclonal antibody is prepared. The prokaryotic expression of His-INMAP fusion protein is induced at 16℃ by adding 0.1 mmol/L isopropyl-β-D-thiogalactoside(IPTG),which is identified by SDS-PAGE analysis and western blotting assay. The inclusion body is treated with4.0 mol/L urea to obtain soluble His-INMAP fusion protein antibody,fusion protein is purified using Ni Sepharose High Performance,and then the protein concentration and purity are detected. The purified protein antibody is injected into 4 Balb/c mice,then blood samples are collected from their hearts,and the anti- serum is isolated. The specificity of polyclonal anti- INMAP antibodies in unpurified and purified prokaryotically expressed products are analysed. In addition,the expression difference between normal liver cell L-02 and 5 hepatoma cell(PLC,Hep G2,SUN449,SMMC-7721 and BEL-7402) is determined by western blotting assay. The results show that His-INMAP fusion protein mainly exists in insoluble inclusion bodies. Soluble protein is obtained with 4.0 mol/L urea treatment to solubilise inclusion bodies. Highly protein purity(94.1%) is harvested after purification. The polyclonal anti-INMAP antibody can bind antigen specifically. Moreover,INMAP is found existing polymorphically in hepatoma cells and its gene expression is down-regulated significantly in all tested hepatoma cells except PLC cell. Obviously,in this study the anti-INMAP polyclonal antibody is of high specificity,which lays a foundation of further study of INMAP functions.
【Fund】: 《科技导报》博士生创新研究资助计划项目(kjdb2012003);; 北京市自然科学基金项目(5122017);; 北京师范大学抗性基因资源与分子发育北京市重点实验室开放基金项目(201204 201307);; 北京师范大学细胞增殖与调控生物学教育部重点实验室开放基金项目(201001)
【CateGory Index】: R735.7;Q78
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 LIU Ping,ZHANG Yun,ZHENG Xi-bang,LI Gong-he,CEN Xiao-mei,YUE Lei-lei,ZONG Zi-jie,LU Sheng-sheng,LU Ke-huan,ZHANG Ming(College of Animal Science and Technology,Guangxi University,Nanning 530005);Preparation of Polyclonal Anti-Sox2 Antibody in Capra hircus[J];Scientia Agricultura Sinica;2012-01
【Secondary Citations】
Chinese Journal Full-text Database 3 Hits
1 ZHANG Qi1,2,XIN Zhenhua1,2,CAI Yinfeng2,GAO Yunxia2,SHA Hongying2,3,LIU Siguo2,CHEN Jianquan2,ZHANG Aimin1,2,CHENG Guoxiang1,2(1.School of Life Science and Medical Engineering,Tongji University,Shanghai 200300;2.Shanghai Transgenic Research Center,Shanghai 201203;3.Huashan Hospital,Fudan University,Shanghai 200040);Cloning,expression and purification of human Sox2 gene[J];Journal of Huazhong Normal University(Natural Sciences);2008-01
2 ZHENG Xi-bang,YANG Zhao-hai,LIU Ping,ZHAO Hua,LU Sheng-sheng,LU Ke-huan,BI Fang-fang (College of Animal Science and Technology,Guangxi University,Nanning 530005,China);Cloning of Capra hircus Sox2,and Its Expression in Escherichia coli and Purification of GST-Sox2 Fusion Protein[J];Chinese Journal of Animal and Veterinary Sciences;2009-10
3 WANG Chun-hong,ZHOU Xiu-mei,LIU Hui,LI Jiang,QIAN Qi-jun(Xinyuan Institute of Medicine and Biotechnology,School of Life Science,Zhejiang Sci-TechUniversity,Hangzhou 310018,China);Cloning Mouse sox2 Gene and Constructing Its Prokaryotic Expression Vector[J];Journal of Zhejiang Sci-Tech University;2010-02
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