| 摘要: |
| CREB(cAMP response element binding protein)是一种真核生物中广泛存在的调控因子。为了探讨企鹅珍珠贝(Pteria penguin)Creb基因的序列和表达特征, 为进一步阐述Creb的生理功能提供科学依据,本研究利用RACE-PCR 技术克隆到企鹅珍珠贝一个Creb基因的全长序列, 分析其理化性质和进化地位; 利用荧光定量PCR分析Creb基因在各组织中的表达特征。结果表明, 该企鹅珍珠贝Creb基因的cDNA 全长为1484 bp, 其中开放阅读框(ORF)为1071bp, 编码356个氨基酸, 5′非编码区为218 bp, 3′非编码区为 195bp, C端包含一个碱性亮氨酸拉链结构(basic region leucin zipper, bZIP)。预测其分子质量为39.49 kD, 等电点为4.43。序列比对显示该企鹅珍珠贝Creb基因与欧洲平牡蛎(Ostrea edulis)和太平洋牡蛎(Crassostrea gigas)的Creb2同源性(identity)最高, 分别为46.3%和46.1%, 故命名为ppCreb2;系统进化树分析结果与传统形态学分类结果相吻合。荧光定量PCR分析显示, ppCreb2在企鹅珍珠贝的各个组织中组成性表达, 其中在足中表达量最高, 鳃中其次。 |
| 关键词: 企鹅珍珠贝 ppCreb2 基因克隆 表达分析 荧光定量PCR |
| DOI：10.11759/hykx20150824001 |
| 分类号: |
| 基金项目:广东海洋大学优秀青年骨干教师培养项目(20140040); 广东海洋大学博士启动基金(E15041); 广东省海洋渔业科技推广专项项目(A201308A11) |
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| Molecular cloning and expression analysis of Creb2 gene from Pteria penguin |
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YU Fei-fei,YU Xiang-yong,PAN Zhen-ni,SONG Na-na,WANG Mei-fang
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| Abstract: |
| In this study, in order to analyze the sequence and expression characteristics of the cAMP response element binding protein (Creb) gene in the Pteria penguin, we characterized the full-length cDNA of the Creb gene from the P. penguin by the rapid amplification of cDNA ends (RACE)-polymerase chain reaction (PCR). We then used real-time PCR to examine the expression of Creb in different tissues. The results show that the full-length cDNA of Creb was 1484 bp, including a 5′?UTR of 218 bp, a 3′ UTR of 195 bp, and an open-reading frame of 1071 bp, which encodes a deduced protein of 356 amino acids. The C-terminal region contained a conserved basic region leucin zipper (bZIP). The predicted molecular weight was 39.49 kD, and the isoelectric point was 4.43. The sequence comparison showed that Creb in the P. penguin (ppCreb2) shares 46.3% and 46.1% sequence identities with Creb2 in Ostrea edulis and Crassostrea gigas, respectively. The phylogenetic analysis results were consistent with traditional taxonomic analysis. The real-time PCR showed that ppCreb2 was constitutively expressed in all studied tissues (mantle, gill, adductor muscle, digestive diverticulum, foot, testis, and ovary), with higher levels in the foot and gill. |
| Key words: Pteria penguin ppCreb2 Gene cloning expression analysis real-time PCR |
