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【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 M5-90 vaccine strain virB2 gene deletion strain. 【Method】 The suicide vector pGEM-7zf-ΔvirB2-sacB was constructed by conventional molecular biology techniques. The transformed Brucella were respectively screened by 100 mg / L ampicillin and 5% sucrose Sensitive screening, access to gene deletion strains. The obtained gene deletion strains were subjected to PCR identification and stability test. 【Results】 The M5-90ΔvirB2 gene deletion strain was successfully constructed, and no mutation was found in the deletion strain within 10 generations. 【Conclusion】 The study laid the foundation for the development of live attenuated Brucella virus attenuated genes.