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

摘要:

摘要：【目的】构建布鲁氏菌virB2基因缺失疫苗，弱化M5-90活苗的毒力。【方法】利用常规分子生物学技术构建同源重组自杀载体pGEM-7zf-ΔvirB2-sacB，电转化布鲁氏菌后分别经过氨苄抗性筛选和蔗糖敏感性筛选，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。

Abstract:

Abstract: [Objective] To construct Brucella vaccine strain M5-90ΔvirB2 deletion mutants with attenuated virulence. [Methods] We constructed the homologous recombination plasmid pGEM-7zf-ΔvirB2-sacB by using traditional molecular biology technology and then obtained M5-90ΔvirB2 strains by ampicillin and sucrose screens after electroporation. The deletion mutants were identified by PCR. Its stability were detected by continuous bacteria culture. [Results] The virB2 deletion mutant strains were successfully constructed. The reverse mutation did not occur within 10 passages. [Conclusion] The results of this study will based on the development of gene deletion attenuated vaccine of Brucella.