Journal of Southern Medical University ›› 2014, Vol. 34 ›› Issue (01): 70-.
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Abstract: Objective To establish a method for gene complementation in Vibrio parahaemolyticus using the plasmid pBAD33.Methods The entire coding region of opaR or aphA was amplified by PCR and cloned into pBAD33. The recombinant plasmidwas transformed into ΔopaR and ΔaphA (the opaR or aphA null mutant strain, respectively) separately to construct thecomplemented mutant strain C-ΔaphA and C-ΔopaR, respectively. RT-PCR was used to verify the transcription of opaR andaphA in the corresponding complemented mutant strains. Primer extension experiments were performed to determine therelative mRNA levels of mfpA (a gene previously characterized to be negatively regulated by AphA and positively by OpaR) inthe wild-type strain, ΔopaR, ΔaphA, C-ΔaphA, and C-ΔopaR. Results opaR and aphA were transcribed in the correspondingcomplemented mutant strains, and their mRNA levels were comparable to those detected in the wild-type strains. ConclusionAmethod has been established for gene complementation in Vibrio parahaemolyticus using the plasmid pBAD33.
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https://www.j-smu.com/EN/Y2014/V34/I01/70