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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]