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ISSN 1672-9145                                                   Acta Biochim Biophys Sin 2004, 36(8): 571–576                                                     CN 31-1940/Q

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

Li-Qing XIAO, Ai-Hua LIU, and Yong-Lian ZHANG*

 

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

 

Abstract        Bin1b is a rat epididymis specific b-defensin which may have fertility related functions in addition to its antimicrobial activity. b-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new b-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of b-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 b-defensin gene family.

 

Key words        Bin1b; epididymis; b-defensin; double-copy protein; polyclonal antiserum

 

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Received: May 28, 2004        Accepted: June 24, 2004

This work was supported by the grants from the National Basic Research Program 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), and Shanghai Science and Technology Development Funding (03JC14080)

*Correspondence author: Tel, 86-21-54921163; E-mail, [email protected]