关键词: 猪, 热休克蛋白, SYBR Green I, 荧光定量PCR, 运输应激

Abstract: 【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 HSP90 mRNA 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.

Key words: Pigs, Heat shock protein, SYBR Green I, FQ-PCR, Transport stress