中国兽医科学2024，Vol.54Issue(1)：34-40,7.DOI:10.16656/j.issn.1673-4696.2023.0217
利用mCherry荧光蛋白报告布鲁氏菌VirB启动子体外活性质粒的构建及应用
Construction and application of in vitro active plasmids of the Brucella VirB promoter were reported using mCherry fluorescent protein
摘要
Abstract
Brucellosis is a zoonotic disease caused by Brucella infection,which has serious harm to the development of breeding industry and human public health and safety.Type Ⅳ secretion systems(T4SS)encoded by VirB are important virulence factors of Brucella.After Brucella infects cells,phagosome acidification is confirmed to be the main signal for VirB to initiate T4SS.Using mCherry red fluorescent protein as a reporter gene,a plasmid that can detect VirB promoter activity in an acidic environment in vitro is constructed,which is important for studying the expression of Brucella viru-lence genes.Plasmid pBE-1-VirB-mCherry was constructed with pBE-1 plasmid as backbone and transferred into Brucella S2 strain by electroporation with pBE-1-ptrc-mCherry plasmid as positive control.Recom-binant strains S2(pBE-1-ptrc-mCherry)and S2(pBE-1-VirB-mCherry)were constructed.Recombinant strain S2(pBE-1-VirB-mCherry)was induced in acidic environment in vitro.The results showed that mCherry fluorescent protein could accurately report the activity of VirB promoter.In conculsion that of a plasmid was constructed to report the activity of Brucella VirB promoter using mCherry fluorescent protein,and it was verified that mCherry fluorescent protein could accurately reflect the activity of Brucella VirB promoter in vitro,which laid a foundation for studying Brucella virulence genes and re-vealing the pathogenic mechanism of its type Ⅳ secretion system.关键词
mCherry荧光蛋白/布鲁氏菌/VirB启动子/Ⅳ型分泌系统Key words
mCherry fluorescent protein/Brucella/VirB promoter/type Ⅳ secretion system分类
农业科技引用本文复制引用
陈彪,侯桂芳,秦建华,王振华,鲍佳鹏,冯紫佳,杨晓彤,孙悦,许行,郑可,许玉静..利用mCherry荧光蛋白报告布鲁氏菌VirB启动子体外活性质粒的构建及应用[J].中国兽医科学,2024,54(1):34-40,7.基金项目
河北省现代农业产业技术体系第三期奶牛创新团队建设项目(HBCT2023180201) （HBCT2023180201）
石家庄圣博生物科技有限公司研发基金项目(SBSW202201) （SBSW202201）
