| An Effective Method for Raising Antisera Against β-defensins： Double-copy Protein Expression of mBinlb in E. coli |
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| 作者姓名： | Xiao LQ Liu AH Zhang YL |
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| 作者单位： | StateKeyLaboratoryofMolecularBiology,InstituteofBiochemist~＂andCellBiology,ShanghaiInstitutesforBiologicalSciences,ChineseAcademyofSciences,GraduateSchooloftheChineseAcademyofSciences,Shanghai200031,China |
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| 摘 要： | Binlb 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 mBin 1 b 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 Bin 1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.
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| 关 键 词： | 附睾 多克隆抗血清 蛋白质 表达 大肠杆菌？A抗血清？A多克隆抗血清 大肠杆菌 蛋白质 表达 附睾 |
An effective method for raising antisera against beta-defensins: double-copy protein expression of mBin1b in E. coli |
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| Xiao LQ,Liu AH,Zhang YL.An effective method for raising antisera against beta-defensins: double-copy protein expression of mBin1b in E. coli[J].Acta Biochimica et Biophysica Sinica,2004,36(8):571-576. |
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| Authors: | Xiao Li-Qing Liu Ai-Hua Zhang Yong-Lian |
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| Institution: | 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. |
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| Abstract: | 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. |
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