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副溶血弧菌是广泛存在于近海区域,盐湖和海产品中的食源性致病菌,会引起大规模的食物中毒。TLH(不耐热溶血毒素)和TDH(耐热直接溶血毒素)是副溶血弧菌最主要的毒力基因,通过比较毒力基因的表达量可以间接比较同种菌株在不同应激条件下以及不同菌株之间的毒力差异。本文以在不同条件下培养的3株Vp为材料,分别提取其总RNA,以16S rRNA为内标基因,运用荧光定量PCR技术检测副溶血弧菌TLH和TDH基因在不同应激条件下的表达差异。结果表明:不同菌株和同种菌株在不同应激条件下tlh、tdh基因表达差异均显著;tlh的最适表达条件分别为5%盐度和20°C;tdh的最适表达条件分别为1%盐度和25°C。运用SPSS软件对实验结果进行统计学分析表明:菌株对tlh表达的影响大于盐度大于温度;菌株对tdh表达的影响大于温度大于盐度。
Vibrio parahaemolyticus is a food-borne pathogen that is widespread in the open sea, salt lakes and seafood and causes large-scale food poisoning. TLH (thermotolerant hemolysin) and TDH (thermotolerans hemolysin) are the most important virulence genes of V. parahaemolyticus. By comparing the expression levels of virulence genes, we can indirectly compare the expression of virulence genes of different strains under different stress conditions and Differences in virulence between different strains. In this paper, three strains of Vp cultured under different conditions were extracted from the total RNA, 16S rRNA as an internal standard gene, the use of fluorescent quantitative PCR detection of Vibrio parahaemolyticus TLH and TDH gene under different stress conditions difference. The results showed that there were significant differences in tlh and tdh gene expression among different strains and isolates under different stress conditions. The optimum conditions for tlh expression were 5% salinity and 20 ° C, respectively, and the optimum conditions for tdh expression were 1 % Salinity and 25 ° C. The statistical analysis of experimental results using SPSS software showed that: the strain of tlh expression greater than the salinity is greater than the temperature; strain tdh expression greater than the temperature is greater than the salinity.