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荧光定量PCR方法分析皱纹盘鲍HSP70在温度胁迫下的表达
程培周1,2, 刘 晓1, 张国范1, 高其康3
1.中国科学院海洋研究所;2.中国科学院研究生院;3.浙江大学分析测试中心
摘要:
根据皱纹盘鲍(Haliotis discus hannai Ino)HSP70和Actin mRNA序列,设计合成引物,建立了皱纹盘鲍HSP70的荧光定量PCR技术平台,并用于检测皱纹盘鲍在热激处理后不同恢复时间HSP70的转录水平变 化。结果表明在热诱导后,肌肉组织HSP70的表达呈时间依赖性,存在先升高后降低的趋势,在热诱导后2h,肌肉HSP70基因的表达量明显升高,是对照 组(0h)的15倍,随后HSP70的表达量继续升高,在诱导后12h达到最高值,是对照组的21倍。然后表达量开始回落,至热诱导后96h,HSP70 表达量降至初始值,表现出一种瞬时表达的趋势。
关键词:  皱纹盘鲍(Haliotis discus hannai Ino)  热激蛋白70  荧光定量PCR
DOI：
分类号:
基金项目:国家863计划项目(2003AA603023,2004AA626070);国家自然科学基金项目(30371117)
Real-time PCR analysis of HSP70 gene expression in response to heat shock
Abstract:
The transcription changes of abalone HSP mRNA were studied during the heat stressing. According to the specific sequence of HSP70 and ACTIN mRNA, the primers were designed and synthesized. By means of established fluorescence quantitative RT-PCR method, the transcription changes of the HSP70 mRNA and the HSP70 mRNA in muscle of transport stressed abalone were studied after 2, 12, 24, 72 and 96 h after heat shock. The volume ratios were calculated as the HSP70 relative expression values. In muscle, levels of HSP70 were significantly increased compared to untreated control abalone, at 2 h of recovery following the heat shock. HSP70 levels reached the peak, at 12 h of recovery. After that, the HSP70 levels dropped till 96 h, when HSP70 returned to control levels.
Key words:  abalone Haliotis discus hannai Ino.  Heat shock protein 70  FQ-PCR
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