半定量RT-PCR方法检测热应激对鲫鱼肝脏中HSP70 mRNA含量的影响

Semi-quantitative RT-PCR Method of Detection of Heat Stress on Fish Liver Content of HSP70 mRNA

依据GenBank已公布的鲫鱼热激蛋白70(Heat shock protein70,HSP70)和beta-actin mRNA序列,分别设计两对引物,应用半定量RT-PCR方法检测应激处理后不同恢复时期鲫鱼(Carassius auratus)肝脏中HSP70 mRNA的表达情况。结果表明,诱导后肝脏组织中HSP70 mRNA的表达量呈现时间依赖性,存在先升高后回落的趋势。在诱导后1h即有所升高,在4h时达到最高值,之后逐渐降低,至48h,表达量降至最初值。研究结果为鲫鱼HSP70的研究提供了基础数据,也为将HSP70用作鱼类生理状态的诊断提供了理论依据。

This research investigated the variation of HSP70 mRNA content in the liver of Carassius auratus at different time lapses after heat stress processing by using semiquantitative RT-PCR.Two primers were designed for this study according to the sequences of Carassius auratus＇s heat shock protein70（HSP70） and beta-actin mRNA published by GenBank.The results show that the HSP70 expression in liver tissue is time-dependent by showing an increase-to-decline trend.The expression starts to increase 1 hour after inducing and reaches a maximum 4 h later. Subsequently,it begins to decline continuously and eventually returns to its initial value until 48 h later.