An Effective Method for Raising Antisera Against β-defensins: Double-copy Protein Expression of mBin1b in E. coli  

作　　者：Li-Qing XIAO Ai-Hua LIU Yong-Lian ZHANG 

机构地区：[1]State Key Laboratory of Molecular Biology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Graduate School of the Chinese Academy of Sciences,Shanghai 200031,China

出　　处：《Acta Biochimica et Biophysica Sinica》2004年第8期571-576,共6页生物化学与生物物理学报（英文版）

基　　金：supported by the grants from the National Basic ResearchProgram of China(No.G1999055901);Director’s Initiating Funding of the Chinese Academy of Sciences;the CAS Knowledge Creative Program(KSCX2-SW-201);the National Natural Science Foundation of China(30230190);Shanghai Science and Technology Development Funding(O3JC14080)

摘　　要：Bin1b is a rat epididymis specific β-defensin which may have fertility related functions in addition to its antimicrobial activity. β-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new β-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of β-defensins make them very difficult to express in prokaryotic system. Here we adopted a double-copy protein expression scheme based on which not only the mBin1b protein was successfully expressed but also the immunity of the antigen was enhanced. The validity of the antisera was verified by using Western blotting and immunohistochemical analyses. It will be a useful tool for deeply investigating the roles of Bin1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.Bin1b is a rat epididymis specific β-defensin which may have fertility related functions in addition to its antimicrobial activity. β-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new β-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of β-defensins make them very difficult to express in prokaryotic system. Here we adopted a double-copy protein expression scheme based on which not only the mBin1b protein was successfully expressed but also the immunity of the antigen was enhanced. The validity of the antisera was verified by using Western blotting and immunohistochemical analyses. It will be a useful tool for deeply investigating the roles of Bin1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.

关 键 词：Bin1b epididymis Β-DEFENSIN double-copy protein polyclonal antiserum 

分 类 号：Q51[生物学—生物化学] Q78