中文摘要：

【目的】探讨应激时热休克蛋白mRNA转录水平的变化。【方法】根据HSP90、HSP10和GAPDH mRNA序列，设计并合成引物，将PCR扩增的基因片段克隆到pGEM-T载体，重组质粒经筛选、鉴定，体外转录出RNA，梯度稀释作为阳性模板，用于标准曲线的制定和样品检测，建立了一步法实时荧光定量RT-PCR技术平台，并用于检测长途运输猪心脏和肝脏中HSP70及HSP90 mRNA转录水平的改变。【结果】经1、2、4、6和10h的运输应激后，组织中HSP70 mRNA的转录水平随着运输应激时间的延长一直呈现出上升的趋势，运输应激到10h时HSP70 mRNA的转录水平升到最高，肝脏和心脏组织中HSP70 mRNA的转录水平分别为对照组的2．5和4．1倍；HSP70 mRNA的转录水平则随着运输应激时间的延长呈下降趋势。【结论】运输应激可以影响HSP mRNA转录水平的变化，并且对不同家族的HSP mRNA转录水平的影响不同；HSP70 mRNA的转录水平有望成为猪运输应激的判定指标之一。

英文摘要：

[ Objective ] To study the transcription changes of the HSP mRNA during the heat stressing. [ Method ] According to the specific sequence of HSP90, HSP70 and GAPDH mRNA, the primers were designed and synthesized. The fragments generated by RT-PCR were cloned into the pGEM-T vector. The positive recombinant plasmids were used to transcript RNA in vitro and the RNA were used as standard quantitative template to make the standard curve and established fluorescence quantitative RT-PCR method. By means of established fluorescence quantitative RT-PCR method, the transcription changes of the HSP70 mRNA and the HSP90 mRNA in livers and hearts of transport stressed pigs were studied after 1 h, 2 h, 4 h, 6 h and 10 h transportation. [ Result ] The level of HSP70 mRNA transcription increased continuously from the beginning of transportation. The inductions of HSP70 mRNA transcription in liver and heart of 10-hour-transport stressed pigs were 2.5 and 4.1 times higher than those of the un-transport stressed pigs （P〈0.01）. However, the transcription levels of HSPaomRNA in livers and hearts decreased with transport stress. [Conclusion] Transport stress can affect the transcription of HSPs mRNA, and the effect is different between HSPs families. Furthermore, the concentration of HSP70 mRNA increased continuously from the beginning of transport stress, might be a hallmark to determine stress.