title: Investigation of Reference Genes in Vibrio parahaemolyticus for Gene Expression Analysis Using Quantitative RT-PCR.
abstract: Vibrio parahaemolyticus is a significant human pathogen capable of causing foodborne gastroenteritis associated with the consumption of contaminated raw or undercooked seafood. Quantitative RT-PCR (qRT-PCR) is a useful tool for studying gene expression in V. parahaemolyticus to characterize its virulence factors and understand the effect of environmental conditions on its pathogenicity. However, there is not a stable gene in V. parahaemolyticus that has been identified for use as a reference gene for qRT-PCR. This study evaluated the stability of 6 reference genes (16S rRNA, recA, rpoS, pvsA, pvuA, and gapdh) in 5 V. parahaemolyticus strains (O3:K6-clinical strain-tdh+, ATCC33846-tdh+, ATCC33847-tdh+, ATCC17802-trh+, and F13-environmental strain-tdh+) cultured at 4 different temperatures (15, 25, 37 and 42°C). Stability values were calculated using GeNorm, NormFinder, BestKeeper, and Delta CT algorithms. The results indicated that recA was the most stably expressed gene in the V. parahaemolyticus strains cultured at different temperatures. This study examined multiple V. parahaemolyticus strains and growth temperatures, hence the finding provided stronger evidence that recA can be used as a reference gene for gene expression studies in V. parahaemolyticus.
authors.affiliation: College of Food Science and Technology, Shanghai Ocean University, Shanghai, People's Republic of China.|Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, Shanghai 201306, China.
authors.forename: Yue-Jiao
authors.initials: YJ
authors.lastname: Ma
authors.affiliation: College of Food Science and Technology, Shanghai Ocean University, Shanghai, People's Republic of China.|Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, Shanghai 201306, China.
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Sun
authors.affiliation: Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Shanghai Ocean University, Ministry of Education, Shanghai, People's Republic of China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Xu
authors.affiliation: College of Food Science and Technology, Shanghai Ocean University, Shanghai, People's Republic of China.|Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, Shanghai 201306, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: College of Food Science and Technology, Shanghai Ocean University, Shanghai, People's Republic of China.|Shanghai Engineering Research Center of Aquatic Product Processing & Preservation, Shanghai 201306, China.
authors.forename: Ying-Jie
authors.initials: YJ
authors.lastname: Pan
authors.affiliation: Residue Chemistry and Predictive Microbiology Research Unit, Eastern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Wyndmoor, PA, 19038, United States of America.
authors.forename: Cheng-An
authors.initials: CA
authors.lastname: Hwang
authors.affiliation: The Pathogenic Microbiology Laboratory, School of Food and Agriculture, University of Maine, Orono, ME, 04469-5735, United States of America.|Produce Safety and Microbiology Research Unit, Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, Albany, CA, 94710, United States of America.
authors.forename: Vivian C H
authors.initials: VC
authors.lastname: Wu
mesh_terms: D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012984:Software; D013696:Temperature; D014736:Vibrio parahaemolyticus / Q000235:genetics* / Q000254:growth & development