半定量RT-PCR检测副溶血弧菌tlh基因的表达差异

Expression of tlh gene in Vibrio parahaemolyticus:semi-quantitative reverse transcription polymerase chain reaction analysis assessment

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是副溶血弧菌最主要的毒力因子之一,通过比较tlh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三株Vp为材料,分别提取其总RNA,以16SrRNA为内标基因,运用半定量反转录—多聚酶链反应(RT-PCR)检测副溶血弧菌不耐热溶血毒素(tlh)基因在不同应激条件下的表达差异。结果表明,在5%的盐度下,三株Vp的tlh基因的表达量都高于其他盐浓度;25℃下,tlh的表达量高于在其他温度条件下;在三株Vp中,ATCC33846中tlhmRNA的表达量最低,ATCC33847中的表达量最高。

Vibrio parahaemolyticus is a foodborne pathogenic bacteria which could be widely found in coastal areas,salt lakes and seafoods, it could cause large-scale food poisoning. TLH is one of the most important virulence factors of Vp, we can compare the virulence of strains which in different stress conditions, as well as differences strains by comparing the expression levels of tlh indirectly. In this paper, three kinds of Vp cultured under different conditions,total RNA were extracted from them, 16S rRNA as an internal standard gene, using semi - quantitative reverse transcription polymerase chain reaction （ RT- PCR） detection of Vp＇ s thermolabile hemolysin toxin（tlh） gene expression under different conditions.The results showed that under 5% of the salinity, tlh gene expression level of the three Vp strains were higher than that under other salt.Under 25℃, tlh gene expression level was higher than that under other temperature conditions.Between the three Vp strains, tlh mRNA expression level of ATCC33846 was lowest, tlh mRNA expression level of ATCC33847 was highest.