布鲁氏菌M5-90疫苗株virB2基因缺失株的构建及鉴定  

Construction and identification of virB2 deletion mutants of Brucella vaccine strain M5-90

作　　者：李臻[1] 张红星[2] 唐利燕[1] 陈创夫[1] 王远志[1] 

机构地区：[1]石河子大学生命科学学院,石河子大学动物科技学院,石河子大学医学院,石河子832003 [2]安阳市中医院,安阳455000

出　　处：《微生物学报》2010年第12期1677-1680,共4页Acta Microbiologica Sinica

基　　金：国家“973项目”(2010CB30203);国际科技合作项目(2006DFA33740);国家自然科学基金项目(30760187,30800813)~~

摘　　要：【目的】构建布鲁氏菌M5-90疫苗株virB2基因缺失株。【方法】利用常规分子生物学技术构建自杀载体pGEM-7zf-ΔvirB2-sacB,通过同源重组的方法,将电转化后的布鲁氏菌分别经100 mg/L氨苄抗性筛选和5%蔗糖敏感性筛选,获得基因缺失株。对获得的基因缺失株进行PCR鉴定和稳定性检测。【结果】成功构建M5-90ΔvirB2基因缺失株,并且该缺失株在10代以内未发生回复突变。【结论】为研发新型布鲁氏菌弱毒基因缺失活苗奠定基础。[Objective]To construct Brucella vaccine strain M5-90 ΔvirB2 mutants.[Methods]Suicide plasmid pGEM-7zf-ΔvirB2-sacB was constructed by traditional molecular biology technology.Through the method of homologous recombination,we screened Brucella gene deletion mutant by 100 mg /L ampicillin resistance screening and 5% sugar sensitivity screening after electroporation.The M5-90 ΔvirB2 mutant was identified by PCR.Its stability was detected by continuous bacteria culture.[Results ] The virB2 deletion mutant strain was successfully constructed.The reverse mutation did not occur within 10 passages.[Conclusion]The results of this study will be based on developing the new attenuated vaccine of Brucella gene deletion mutants.

关 键 词：布鲁氏菌 virB2基因 基因缺失株 

分 类 号：R392[医药卫生—免疫学]