An effective method for raising antisera against beta-defensins: double-copy protein expression of mBin1b in E. coli.
Bin1b is a rat epididymis specific beta-defensin which may have fertility related functions in addition to its antimicrobial activity. beta-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new beta-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of beta-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 beta-defensin gene family.
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.
Li-Qing
LQ
Xiao
Ai-Hua
AH
Liu
Yong-Lian
YL
Zhang
D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D004822:Epididymis / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage; D015870:Gene Expression; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D023083:beta-Defensins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D018076:DNA, Complementary; D011994:Recombinant Proteins; D023083:beta-Defensins