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。
Scavenger decapping enzyme （DcpS） is a kind ofmRNA decapping enzyme, being tightly associated to the regulation of gene expression, respiration, activity of motor neurons, cancer and various stress responses. In this study, the full length of PmDcpS gene was obtained using rapid amplification of cDNA ends （RACE） technology, and expression of PmDcpS in different tissues was tested by real-time PCR. Results showed that the total length ofPmDcpS gene was 1 034 bp, including a 5＇-terminal non-coding region （UTR） of 25 bp, a 3＇ UTR of 34 bp with 29 bp poly A, and an open reading frame （ORF） of 975 bp which encodes 324 amino acids. The predicted molecular weight was 37.94 kD, isoelectric point was 6.12. Sequence analysis found that PmDcpS has the typical DcpS structure domain. Multiple sequence alignment showed that DcpS was highly conservative among species, and thehomology of PmDcpS to Crassostrea gigas was 65.6%. Real-time PCR data revealed that PmDcpS could be expressed in all of the tested tissues, including hemocytes, adductor muscle, mantle edge, mantle central,gill, gonads, hepatopancreas, with the higher expression in mantle central and hepatopancreas. This study could provide basis for the further study of the biological function of PmDcpS in the shellfish.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第8期3210-3218,共9页
Genomics and Applied Biology
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
