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从鲫鱼的肠道内提取总RNA,运用RT-PCR扩增ghrelin基因并测序,用生物信息学软件预测其结构,并构建系统进化树,探讨了鲫鱼ghrelin基因的生物学功能,为进一步研究该基因对鲫鱼摄食的影响奠定基础。结果显示,获得了长490bp的ghrelin基因(GenBank登录号为HM567312),其序列含有1个312bp的完整开放阅读框,编码103个氨基酸;ghrelin蛋白分子质量为11 519.48u,等电点为5.20,半衰期为1.20h,不稳定系数为31.29,总平均亲水性为0.727,疏水性为-2.900～2.856;1～26位氨基酸可能是信号肽序列;在ghrelin的分子序列中存在44处α螺旋富集区域,8处β折叠区域,集中于成熟肽部分,有43处无规则卷曲。系统进化树分析显示鲫鱼ghrelin基因与金鱼的进化距离最近。
The ghrelin gene was amplified by RT-PCR and sequenced. The structure of ghrelin gene was predicted by bioinformatics software. The phylogenetic tree was constructed to investigate the biological function of ghrelin gene in crucian carp. The crucian carp feeding basis. The results showed that the ghrelin gene (GenBank accession number HM567312) with a length of 490 bp was obtained. The sequence contained a complete open reading frame of 312 bp and encoded a protein of 103 amino acids. The ghrelin protein had a molecular mass of 11 519.48 u and an isoelectric point of 5.20. The half-life was 1.20h, the instability coefficient was 31.29, the total average hydrophilicity was 0.727, the hydrophobicity was -2.900 ~ 2.856; the amino acids 1 ~ 26 were probably signal peptide sequences; there were 44 α-helix rich in the molecular sequence of ghrelin Set of regions, 8 β-sheet folding regions, concentrated in the mature peptide fraction, 43 irregularly curled. Phylogenetic tree analysis showed that the ghrelin gene in crucian carp was closest to goldfish.