Pathogenic Vibrio campbellii HY01, a major causative agent of luminous vibriosis in shrimp, was used as a representative strain to investigate the effects of temperature and salinity on bacterial survival, virulence, and gene expression. The strain HY01 was cultured in Luria-Bertani (LB) broth supplemented with artificial seawater at 30 (optimal) and 60 ppt salinity (salt stress condition), and incubated at two temperatures: 30 (optimal temperature) and 35 (warm temperature) degrees Celsius. During the mid-exponential growth phase, bacterial cells were harvested, and total RNA was extracted. RNA sequencing and data analysis were then performed.