缢蛏热休克转录因子1 (HSF1) 基因克隆、组织表达及功能

Cloning and tissue expression of heat shock transcription factor 1 (HSF1) gene of Sinonovacula constricta

  • 摘要: 热休克转录因子1 (HSF1) 是一种广泛存在于真核生物中的调控因子,在生物体内具有多种生理功能。为了探究海洋滩涂生物在耐受高温过程中HSF1发挥的作用和分子机制,进一步阐述HSF1的生理功能,文章以缢蛏 (Sinonovacula constricta) 为实验材料,利用RACE技术克隆HSF1基因。结果显示,缢蛏HSF1基因的cDNA全长2 026 bp,开放阅读框 (ORF) 长1 707 bp,编码568个氨基酸,5'非编码区 (UTR) 长196 bp,3' UTR长123 bp。氨基酸序列比对及系统进化树结果显示缢蛏HSF1基因与美洲牡蛎 (Crassostrea virginica) 的亲缘关系最为接近,其系统进化分析与传统的形态学分类相吻合。荧光定量PCR结果显示,HSF1基因在各个组织均有表达,其中在外套膜中的相对表达量最高,其次是鳃、肝胰腺和水管,在斧足和性腺组织的相对表达量较低 (P<0.05)。荧光定量结果显示HSF1基因在急性温度胁迫后第9小时表达量较高。推测HSF1基因参与缢蛏的热应激过程,并起到一定作用。

     

    Abstract: The heat shock transcription factor 1 (HSF1) is a regulatory factor widely present in eukaryotes and has many physiological functions in vivo. In order to explore the role and molecular mechanism of HSF1 in the process of high temperature tolerance, and further elaborate its physiological function, we cloned the HSF1 gene of Sinonovacula constricta by RACE technology. The results show that the full-length cDNA of S. constricta HSF1 gene was 2 026 bp; the open reading frame was 1 707 bp in length, encoding 568 amino acids; the 5' non-coding region was 196 bp in length; the 3' non-coding region was 123 bp in length. Amino acid sequence alignment and phylogenetic tree results show that HSF1 gene was the closest to the Crassostrea virginica, and its phylogenetic analysis was consistent with traditional morphological classification. The results of real-time PCR show that HSF1 gene was expressed in all tissues, and the relative expression of mantle membrane was the highest, followed by sputum, hepatopancreas and water tube, and the relative expressions in axillary and gonadal tissues were low.

     

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