鳜三种生长抑素与两种受体cDNA全长克隆及组织表达分析-动物遗传育种与繁殖专业论文.docxVIP

上海海 上海海洋大学硕士学位论文 II II 7 个转膜区（TMD）、3 个细胞外袢（ECLs）、4 个细胞内袢（ICLs）和 C 末端区域。 NJ 系统进化树结果显示，鳜 SSTR2 和 SSTR3 分别形成相对独立的分支，两者氨 基酸序列相似度为 51.2%，表明它们是由不同基因编码而成。利用实时荧光定量 RT-PCR 技术检测了 SSTR2 和 SSTR3 mRNA 的组织表达特征，它们均在多种组织 中广泛表达。其中，SSTR2 mRNA 在肝脏中表达量最高，其次是脑、脾脏、性腺 和幽门垂，心脏、食道、胃、肠道中表达量较低。SSTR3 mRNA 在胃中表达量最 高，其次是脑、食道、肾和性腺，鳃和幽门垂中表达量较低。SSTR2 和 SSTR3 mRNA 均在脑中表达，与三种 PSS 均在脑中表达一致，推测鳜 SS 主要参与脑中的神经和 激素调控。SSTR2 mRNA 在肝脏中表达量最高，SSTR3 mRNA 在胃中表达量最高, 推测 SS 在肝脏中可能主要通过与 SSTR2 结合，参与 GH-IGF 生长轴调控；在胃中 可能主要通过与 SSTR3 受体结合，参与消化作用调节。 关键词：鳜；生长抑素；生长抑素受体；RACE；组织表达；原位杂交 PAGE PAGE III Cloning and expression analysis of three somatostatins and two somatostatin receptors in Siniperca chuatsi ABSTRACT Somatostatins (SS) play important physiological functions in vertebrates growth and development, which belong to a multiple gene family. Combining with the somatostatin receptor (SSTR), they could regulate individual development and growth, though the growth hormone - insulin-like growth factor - I (GH-IGF-Ⅰ) axis at multiple levels. In this study, the complete cDNA sequences of three distinct preprosomatostatins (PSSⅠ, PSSⅡand PSSⅢ) were isolated from Siniperca chuatsi brain and cloned by means of rapid amplification of cDNA ends (RACE). The full length of PSSⅠ cDNA was 647 bp, which contained 369 bp open reading frame (ORF), and encoded 122 amino acids with a signal peptides of 26 amino acids, it contained the conservative SS-14 sequence at the C-terminal extremity, which is identical to that of other vertebrates. The full length of PSSⅡ cDNA was 655 bp, which contained 387 bp open reading frame (ORF), encoded 128 amino acids with a signal peptides of 25 amino acids, it contained [Tyr7, Gly10]SS-14 at the C-terminal extremity. The full length of PSSⅢ cDNA was 823 bp, which contained 333 bp open reading frame (ORF), encoded 110 amino acids with a signal peptides of 21 amino acids, it contained [Pro2]SS-14 at its C-terminal extremity. PSS mRNA expression in different adult tissues were analyzed by RT-PCR technique,