Qin Li1, Zhiyun Peng, Xinpeng Chen, Xiaohong Sun, Yingjie Pan, Yong Zhao. 1. College of Food Science and Technology, Shanghai Ocean University, Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai Engineering Research Center of Aquatic-Product Processing & Preservation, Shanghai 201306, China. lqapple1988@yahoo.com.cn
Abstract
OBJECTIVE: We selected reference genes for virulence gene expression of Vibrio parahaemolyticus under different environmental conditions. METHOD: Using qRT-PCR, we evaluated the expression stability of four housekeeping genes (GAPDH, pvuA, pvsA and rpoS) of V. parahaemolyticus cultivated separately in seawater, filtered seawater, shrimp and Tryptone Soya Broth. RESULT: The result shows that all the candidate reference genes could be amplified specifically in qRT-PCR reaction. The expression stability of the four reference genes ranked pvuA (2.906) > pvsA (3.197) > GAPDH (3.746) > rpoS (6.512). Further analysis with geNorm program reveals that the highest stability was observed in pvuA and pvsA. The geometric average score of the two genes was considered as the most appropriate reference gene for normalizing the expression of virulence genes of V. parahaemolyticus. CONCLUSION: The pvuA and pvsA genes could be used as reference genes to study the virulence gene expression of V. parahaemolyticus.
OBJECTIVE: We selected reference genes for virulence gene expression of Vibrio parahaemolyticus under different environmental conditions. METHOD: Using qRT-PCR, we evaluated the expression stability of four housekeeping genes (GAPDH, pvuA, pvsA and rpoS) of V. parahaemolyticus cultivated separately in seawater, filtered seawater, shrimp and Tryptone Soya Broth. RESULT: The result shows that all the candidate reference genes could be amplified specifically in qRT-PCR reaction. The expression stability of the four reference genes ranked pvuA (2.906) > pvsA (3.197) > GAPDH (3.746) > rpoS (6.512). Further analysis with geNorm program reveals that the highest stability was observed in pvuA and pvsA. The geometric average score of the two genes was considered as the most appropriate reference gene for normalizing the expression of virulence genes of V. parahaemolyticus. CONCLUSION: The pvuA and pvsA genes could be used as reference genes to study the virulence gene expression of V. parahaemolyticus.