Polyclonal antibody preparation and expression in liver tissues of transactivated protein 5 of hepatitis C virus nonstructural 5A  

作　　者：Xiao-quan Li1,2,Shu-lin Zhang2,Li-hua Zhong3,Jun Cheng3,Yuan Hong3,Meng-dong Lan2,Xiao-bin Chen3,Cheng-fu Sun31. Department of Pediatrics,the First Affiliated Hospital,Medical School of Xi’an Jiaotong University,Xi’an 710061 2. Department of Infection,the First Affiliated Hospital,Medical School of Xi’an Jiaotong University,Xi’an 710061 3. Institute of Infectious Diseases,Beijing Ditan Hospital,Beijing 100011,China. 

出　　处：《Journal of Pharmaceutical Analysis》2009年第2期99-103,共5页药物分析学报（英文版）

基　　金：supported by the National Natural Science Foundation of China (No.C03011402and30371288)

摘　　要：Objective To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods In Escherichia coli BL21,the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG),and it was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyclonal antibody,with which we studied the function of NS5ATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results Recombinant NS5ATP5 (molecular weight:65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person,and it was detected mainly in the cytoplasm. Conclusion The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.Objective To prepare polyclonal antibody of transactivated protein 5 of hepatitis C virus nonstructural 5A (NA5ATP5) and to explore its expression in the liver tissues. Methods In Escherichia coli BL21,the prokaryotic expression vector pET32a(+)-NS5ATP5 was induced by isopropyl-β-D-thiogalactoside (IPTG),and it was analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. And the purified protein was used to immunize the rabbit to prepare polyclonal antibody,with which we studied the function of NS5ATP5 by determining the different liver tissues with the streptavidin-perosidase (SP) immunohistochemistry method. Results Recombinant NS5ATP5 (molecular weight:65 kD) and polyclonal antibody were successfully prepared. NS5ATP5 expression in the liver of patients with chronic HCV infection was much higher than that of a normal person,and it was detected mainly in the cytoplasm. Conclusion The findings of the expression difference between HCV patients and normal people led to a novel diagnostic marker to detect HCV infection.

关 键 词：NS5ATP5 polyclonal antibody IMMUNOHISTOCHEMISTRY 

分 类 号：R512.63[医药卫生—内科学]