Construction and application of in vitro active plasmids of the Brucella VirB promoter were reported using mCherry fluorescent protein
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.
mCherry fluorescent proteinBrucellaVirB promotertype Ⅳ secretion system
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