摘要
组蛋白H3是组成核小体的基本组分之一。研究表明,组蛋白不仅参与染色质的组装,也可以通过调节基因表达参与调控细胞分裂、凋亡、DNA修复等细胞生命过程。为了探究马氏珠母贝组蛋白H3(Pinctada martensii histone H3,Pm H3)的序列及表达特征,本文运用c DNA末端快速扩增(RACE)技术克隆得到Pm H3的c DNA全长序列,并且应用实时荧光定量PCR技术对Pm H3基因在马氏珠母贝不同组织中的表达进行检测。结果显示,Pm H3基因序列全长627 bp,其中开放阅读框(ORF)为411 bp,编码136个氨基酸,5’UTR为42 bp,3’UTR为174 bp,包含26 bp的poly A。预测其相对分子量为15 388.0 Da,理论等电点为11.27,不稳定系数为47.19,疏水性水平-0.604。多序列比对结果表明H3基因极为保守,在各物种间的相似度达到99%~100%。SMART软件分析发现Pm H3具有一个典型的H3结构域。荧光定量PCR数据结果显示,Pm H3基因在马氏珠母贝外套膜边缘区、外套膜中央区、鳃、血细胞、闭壳肌、足、性腺、肝胰腺八种组织中均有表达,其中在性腺中表达量最高。本研究可为进一步探讨Pm H3基因在贝类中的生物学特性以及组蛋白修饰提供理论依据。
Histone H3 is one of the basic components of nucleosome. Increasing evidence has showed that histone was not only involved in chromatin assembly, but also participated in various biological processes, such as cell division and apoptosis, DNA repair, by regulating gene expression. In order to explore the sequence and expression characteristics of histone H3 in Pinctada martensii, using c DNA End Rapid Amplification(RACE)Cloning technology, we have got the c DNA full length of histone H3 from P. martensii(Pm H3) and analyzed its expression patterns in different tissues by Real-Time PCR technology. Results showed that the total length of Pm H3 gene was 627 bp, including 411 bp of the open reading frame(ORF) which encodes 136 amino acids, a 5' UTR of 42 bp and a 3' UTR of 174 bp with 26 bp poly A. The predicted molecular weight is 15 388.0 Da, and the isoelectric point is 11.27, instability index is 47.19, and grand average of hydropathicity(GRAVY) is-0.604. Multiple sequence alignment results showed Pm H3 gene was highly conservative among species with 99%~100% identities.SMART software analysis showed that Pm H3 has a typical H3 structure domain. Real-time PCR data revealed Pm H3 gene could be detected in all of tested tissues, including mantle central, mantle edge, gill, haemocytes,adductor muscle, foot, gonads, and hepatopancreas, with the highest expression level in the gonads. Our study would not only help to further illustrate the function of Pm H3 gene in the shellfish, but also provide the theory basis to our understanding of the mechanism underlying the histone modification in gene regulation.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2015年第7期1421-1428,共8页
Genomics and Applied Biology
基金
国家自然科学基金项目(31471271)
广西壮族自治区重点实验室基金项目
广西高校重点实验室基金项目共同资助
