南方医科大学学报Issue(1)：70-74,5.
副溶血弧菌基因回补实验方法的建立与应用
Establishment of a method for gene complementation in Vibrio parahaemolyticus
摘要
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 plasmid was transformed into ΔopaR and ΔaphA (the opaR or aphA null mutant strain, respectively) separately to construct the complemented mutant strain C-ΔaphA and C-ΔopaR, respectively. RT-PCR was used to verify the transcription of opaR and aphA in the corresponding complemented mutant strains. Primer extension experiments were performed to determine the relative mRNA levels of mfpA (a gene previously characterized to be negatively regulated by AphA and positively by OpaR) in the wild-type strain, ΔopaR, ΔaphA, C-ΔaphA, and C-ΔopaR. Results opaR and aphA were transcribed in the corresponding complemented mutant strains, and their mRNA levels were comparable to those detected in the wild-type strains. Conclusion A method has been established for gene complementation in Vibrio parahaemolyticus using the plasmid pBAD33.关键词
副溶血弧菌/基因回补/pBAD33/aphA/opaRKey words
Vibrio parahaemolyticus/gene complementation/pBAD33/aphA/opaR引用本文复制引用
陈振鸿,王丽,张义全,冯娇,杨瑞馥,常德,安莉,刘长庭,周冬生..副溶血弧菌基因回补实验方法的建立与应用[J].南方医科大学学报,2014,(1):70-74,5.基金项目
国家自然科学基金（31170127，81200059）；国家重点基础研究发展计划973项目（2014CB744400）Supported by National Natural Science Foundation of China（31170127,81200059） and National Basic Research Program of China（973 program） （）
