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[目的]克隆香鱼热休克转录因子1(Heat shock transcription factor1,HSF1)基因,并探究镉胁迫对其表达的影响。[方法]利用3’RACE和5’RACE技术克隆香鱼HSF1基因c DNA全长,并进行序列分析,通过半定量PCR(RTPCR)检测HSF1在各组织中的表达情况,通过实时荧光定量PCR(qRT-PCR)检测分析镉胁迫后HSF1表达量的变化。[结果]香鱼HSF1基因c DNA序列全长2 127 bp(不含poly A),含一个1 587bp、编码528个氨基酸的开放阅读框(ORF);预测其编码蛋白分子量为57.956 87 k Da,等电点为4.56。与虹鳟进化关系最近。反转录PCR(RT-PCR)显示香鱼HSF1基因在鳃中表达量最低、肌肉中次之,其余组织中基本一致。qRT-PCR结果显示镉胁迫后香鱼肝、心和肌肉中HSF1基因表达量均显著增加,脾、肾和鳃中表达量均显著减少。[结论]HSF1基因参与香鱼镉胁迫应激反应。
[Objective] The aim of the study was to clone the gene of Heat shock transcription factor 1 (HSF1) and explore the effect of cadmium stress on its expression. [Method] The full-length c DNA of HSF1 gene was cloned by 3’RACE and 5 ’RACE techniques and sequenced. The expression of HSF1 in various tissues was detected by semi-quantitative PCR (RTPCR) qRT-PCR) detection of cadmium stress HSF1 expression changes. [Result] The full length cDNA of HSF1 gene from sweetfish was 2 127 bp (without poly A) containing a 1 587 bp open reading frame (ORF) encoding 528 amino acids. The predicted molecular weight of HSF1 gene was 57.956 87 kDa, The isoelectric point is 4.56. Evolution of rainbow trout and the recent relationship. Reverse transcriptase PCR (RT-PCR) showed that the expression level of HSF1 was the lowest in the gill and the second was in the muscle, and the other tissues were basically the same. qRT-PCR results showed that HSF1 gene expression in liver, heart and muscle significantly increased after cadmium stress, but decreased in spleen, kidney and gill. [Conclusion] The HSF1 gene is involved in the stress response of sweet-scented osmanthus to cadmium stress.