【摘要】 从褐菖鲉肝脏中克隆了热休克蛋白HSC70基因,利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)建立HSC70基因的定量检测方法。为检测该方法的可行性,将褐菖鲉分别暴露于石油水溶性成分(water-soluble fraction,WSF)20、60、180μg·L-11 d后,利用real-time PCR及LAMP技术同时测定褐菖鲉肝HSC70 mRNA表达量,两种方法测定结果基本一致,证实LAMP技术可用于褐菖鲉肝HSC70基因的定量。为更细致了解石油WSF影响褐菖鲉肝脏HSC70基因表达的剂量-效应关系,将褐菖鲉分别暴露于25、50、75、100、125、150、175μg·L-1WSF中,5 d后采样,用LAMP技术定量检测HSC70 mRNA。结果表明,HSC70 mRNA表达量在50μg·L-1浓度组即被显著诱导,在75μg·L-1浓度下达到最大值,这说明褐菖鲉肝HSC70基因对石油污染较敏感,有潜力作为海洋石油污染的生物标志物。更多还原

【Abstract】 Heat shock cognate protein(HSC70) gene was cloned from the liver of Sebastiscus marmoratus. A quantitative loop-mediated isothermal amplification(LAMP) method was established to analyze the mRNA level of HSC70 in S. marmoratus. S. marmoratus had been exposed to different concentrations(20, 60, 180 μg·L-1) of the water-soluble fraction(WSF) of crude oil for 1 day. Then, the contents of HSC70 mRNA in the liver of S. marmoratus weremeasured using real-time PCR and LAMP. The change patterns of HSC70 mRNA levels measured by LAMP were consistent with those measured by real-time PCR, indicating that the LAMP method was feasible for quantify HSC70 mRNA. To clearly understand the dose-effect relationship between WSF and HSC70, S. marmoratus had been exposed to WSF of seven different concentrations(25, 50, 75, 100, 125, 150, 175 μg·L-1) for 5 days, and the mRNA levels of HSC70 in liver were examined by LAMP. The results showed that the HSC70 mRNA levels were significantly increased in 50 μg·L-1group and reached the peak in 75 μg·L-1group, indicating that HSC70 in the liver of S. marmoratus was sensitive to WSF, and could be used as a potential biomarker to oil pollution.更多还原