知识与财富的链接
HDAC1作为HDACs家族中重要成员,可使组蛋白去乙酰化进而调节基因表达,在细胞分化和胚胎早期发育中起重要作用。本文利用SMART RACE技术克隆得到泥蚶HDAC1(Tg-HDAC1)基因的cDNA全长序列,并对其内含子进行扩增及不同组织、不同发育时期的定量表达。结果发现:Tg-HDAC1基因的cDNA序列全长为2275 bp,开放阅读框(ORF)1587 bp,编码528个氨基酸;Tg-HDAC1蛋白序列与斑马鱼、鸡、小鼠等的相似性都在80%以上,表明该蛋白氨基酸序列在物种进化过程中具有保守性;在Tg-HDAC1基因中扩增出13个内含子,均存在于开放阅读框中,且都遵循GT-AG原则;荧光定量PCR(qRT-PCR)分析结果显示,Tg-HDAC1基因在成体血液、内脏团、外套膜、鳃、斧足和闭壳肌6个组织中均有表达,而在足中的表达量最高,且与其余5组织中的表达量有极显著差异(P<0.01),说明它对该组织生长具有重要作用。不同发育时期的表达差异结果表明,Tg-HDAC1基因在担轮幼虫期表达量最高,显著高于其他发育时期(P<0.05)。
As an important member of HDACs family, HDAC1 can regulate gene expresstion and play a crucial role in cell differentiation and early embryonic development. Tg-HDAC1 cDNA was cloned by SMART RACE technique and then the bioinformatics, expression analysis, and intron amplification of Tg-HDAC1 were carried out in Tegillarca granosa. The full length of Tg-HDAC1 cDNA was 2 275 bp, containing a complete 1587 bp ORF encoding 528 amino acids. The homologous similarity between the blood clam and other species, such as Danio rerio, Gallus gallus, Mus musculus, was more than 80%, which indicate that HDAC1 is relatively conserved in the evolution. Thirteen introns of Tg-HDAC1 were amplified in ORF, which all of them follow the principle of GT-AG. The results of six tissue-specific expression by real time PCR showed that Tg-HDAC1 gene expressed in all tissues, and the expression of foot were significantly higher than other tissues(P<0.01), which suggest that the gene play an important role in the course of foot growth. The relative expression in different stages revealed that the expression of Tg-HDAC1 gradually increased with the process of the development, and showed the highest in trochophore stage (P<0.05).
