马氏珠母贝DcpS基因的分子特征与表达分析

发布时间：2018-11-21 21:05

【摘要】：清道夫脱帽酶(DcpS)是一种mRNA脱帽酶,与基因表达调控、呼吸、运动神经元活动,癌症和各种应激反应紧密相关。本研究采用cDNA末端快速扩增(RACE)技术克隆获得了马氏珠母贝DcpS(Pm Dcp )基因cDNA全长序列;运用实时荧光定量PCR(Real-time PCR)技术检测Pm DcpS基因在马氏珠母贝不同组织中的表达模式。结果显示,Pm DcpS基因序列全长1 034 bp,其中开放阅读框(ORF)为975 bp,编码324个氨基酸,5'非编码区(5'UTR)长为25 bp,3'非编码区(3'UTR)长为34 bp,包含29 bp的poly A。预测分子量约为37.94 k D,理论等电点为6.12。序列分析发现Pm DcpS具有典型DcpS结构域。多序列比对结果表明物种间DcpS具有较高的保守性,与太平洋牡蛎的相似性为65.6%。实时荧光定量PCR数据分析表明,该Pm DcpS基因在马氏珠母贝性腺、鳃、肝胰腺、血细胞、闭壳肌、外套膜边缘区、外套膜中央区这7种组织中均有表达,其中在外套膜中央区和肝胰腺表达量较高。本研究可为进一步探究Pm DcpS在贝类中的生物学功能提供重要的理论基础和参考价值。
[Abstract]:Scavenger discap enzyme (DcpS) is a mRNA discap enzyme, which is closely related to gene expression regulation, respiration, motor neuron activity, cancer and various stress responses. In this study, the full-length cDNA sequence of DcpS (Pm Dcp) gene of Pinctada martensii was cloned by rapid amplification of (RACE) at the end of cDNA. The expression patterns of Pm DcpS gene in different tissues of Pinctada martensii were detected by real-time fluorescence quantitative PCR (Real-time PCR). The results showed that the total length of, Pm DcpS gene was 1 034 bp,. The open reading frame (ORF) encoded 324 amino acids for 975 bp, and the 5 'non-coding region (5'UTR) was 25 bp,3' non-coding region (3'UTR). The length of 3'UTR was 34 bp,. Poly A. The predicted molecular weight is about 37.94 KD and the theoretical isoelectric point is 6.12. Sequence analysis showed that Pm DcpS had a typical DcpS domain. The results of multiple sequence alignment showed that DcpS among species was highly conserved, and the similarity with Pacific oyster was 65.6. Real-time fluorescence quantitative PCR analysis showed that the Pm DcpS gene was expressed in the gonad, Gill, hepatopancreas, blood cells, obturator muscle, marginal region of coat membrane and central region of coat membrane in seven kinds of tissues, such as gonad, Gill, hepatopancreas, blood cells, adductor muscle, and central region of coat membrane. The expression level was higher in the central region of coat membrane and hepatopancreas. This study can provide important theoretical basis and reference value for further exploring the biological function of Pm DcpS in shellfish.
【作者单位】：广东海洋大学水产学院;
【基金】：国家自然科学基金“马氏珠母贝植核育珠移植耐受免疫相关基因与蛋白质鉴定及关联度研究”(31472306) 广东省科技计划项目“海水珍珠贝功能基因的开发与应用研究”(2012A031100010) 广东省海港建设与渔业产业发展专项”马氏珠母贝抗菌功能多肽的提取与应用研究“(A201608B15)共同资助
【分类号】：Q78;S917.4

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