| 摘要: |
| 采用 RT-PCR 和 RACE 技术克隆了大黄鱼胰岛素样生长因子-Ⅰ(IGF-Ⅰ) cDNA 的全序列, 序列全长 2027bp, 由 57bp 的 5’非翻译区、1409bp 的 3’非翻译区和 561bp 的开放阅读框组成。序列分析表明, 大黄鱼 IGF-Ⅰ编码区包括信号肽、成熟肽(B、C、A、D)和 E 等 6 个区域的 186 个氨基酸, 形成成熟肽时, 信号肽和 E 区被切除, 成熟肽分子量 7.5kDa, 等电点(pI)为 7.76, 成熟肽的 B 区和 A区有 6 个保守的半胱氨酸残基, 形成 3 对二硫键, 起到了稳定 IGF-Ⅰ三维结构的作用。与其它物种的IGF-Ⅰ比对后发现A、 B区域比较保守, C、 D区域保守性较差; E区域的分析结果表明, 大黄鱼IGF-ⅠcDNA序列为Ea-4亚型。 核苷酸同源性分析发现, 大黄鱼IGF-ⅠcDNA与美国红鱼同源性最高, 为99.47%; 与斜带石斑鱼、金头鲷和褐牙鲆的同源性依次降低, 分别为 97.68%、97.50%和 95.90%。 |
| 关键词: 大黄鱼, 胰岛素样生长因子-Ⅰ(IGF-Ⅰ), 克隆 |
| DOI：10.11693/hyhz201201005005 |
| 分类号: |
| 基金项目:国家农业公益性行业科研专项“优质安全大黄鱼养殖产业链技术研究与示范”, 200903029-06 号; 国际科技合作项目“浅海典型渔业生态功能恢复与重建关键技术”, 2009DFB20290 号 |
|
| MOLECULAR CLONING AND SEQUENCE ANALYSIS OF INSULIN-LIKE GROWTH FACTOR ⅠcDNA FROM LARIMICHTHYS CROCEA |
|
CHANG Kang-Mei, ZHENG Xiao-Liang, LIU Hui-Hui, ZHANG Jian-She, WU Chang-Wen
|
|
Marine Science College of Zhejiang Ocean University, Key Laboratory of Zhejiang Provincial Marine Culturing Equipments and Engineering Technology
|
| Abstract: |
| The insulin-like growth factor (IGF-Ⅰ) gene of Larimichthys crocea was cloned from liver by RT-PCR and RACE technique. All the length of the nucleotide sequence was 2027bp, including 5’UTR (57bp), 3’UTR (1409bp) and ORF (561bp). Nucleotide sequence analysis showed that ORF encoded 186 amino acids that was composed of signal peptide, mature peptide (B, C, A, D) and E domain. Signal peptide and E domain were cut off when the peptide was processed. The molecular weight of mature peptide was 7500Da and its pI was 7.76. There were three disulfide bonds formed by six cysteines in B and A domains. The disulfide bonds comparing the amino acid sequence of IGF-Ⅰof L. crocea with those of other species, the sequence conservation of A and B domains is high, while low in C and D domains. E domains in IGF-Ⅰof L. crocea was belonged to IGF-ⅠEa-4 subtype. Analysis of nucleotide sequence showed that IGF-ⅠcDNA of L. crocea is similar to other fishes which shared sequence homology of 99.47% with Sciaenops ocellatus, 97.68% with Epinephelus coioides, 97.50% with Sparus aurata and 95.90% with Paralichthys olivaceus. |
| Key words: Larimichthys crocea, IGF-Ⅰ, Cloning |
