Herpes Simplex Virus Type 1 ICP27 Protein:Its Expression, Purification and Specific Antiserum Production  

作　　者：Lei ZHAO Xiao-ming REN Alan C. ZHENG 

机构地区：[1].State Key Laboratory, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China [2]The Depautment of Animal Science, Beijing University of Agriculture, Beijing 102206, China

出　　处：《Virologica Sinica》2010年第3期199-205,共7页中国病毒学（英文版）

基　　金：The startup fund of the hundred talents program of the Chinese academy of science(20071010- 141);National natural science foundation of China (30870120);Open research fund program of the state key laboratory of virology of China (2007003, 2009007);Hubei province natural science foundation of innovation groups project (2008CDA013);Major state basic research development program (973 Program) of China (2010CB 530105)

摘　　要：Herpes simplex virus type 1 (HSV-1) is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 (ICP27) is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a (+) vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid (Ni 2+ -NTA) affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay (IFA). Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27's biological function during HSV-1 infection.Herpes simplex virus type 1 （HSV-1） is the causative agent of cold sores and other more serious diseases. HSV-1 infected-cell protein 27 （ICP27） is an immediate-early regulatory phosphoprotein homologous to gene products identified in all classes of herpesviruses so far. To raise the antiserum to ICP27 for further characterization of its biological function, the ICP27 gene was cloned into the pET-28a （＋） vector, then ICP27 protein was expressed in E. coli and purified by nickel-nitrilotriacetic acid （Ni^2＋-NTA） affinity resin column, finally the purified protein was used to raise antiserum. Western blot analysis demonstrated that the antiserum recognized the recombinant protein, and the antiserum was able to probe the ICP27 in HSV-1 infected cells with high specificity by immunofluorescence assay （IFA）. Therefore, the specific antiserum will provide a valuable tool for further studies investigating ICP27＇s biological function during HSV-1 infection.

关 键 词：Herpes simplex virus type 1 （HSV-1） Infected-cell protein 27 （ICP27） Recombinant protein ANTISERUM Immunofluorescence assay. 

分 类 号：S432.41[农业科学—植物病理学] Q78[农业科学—农业昆虫与害虫防治]