摘要
从斑马鱼脑组织提取总RNA,应用RT-PCR方法克隆促性腺激素释放激素(GnRH)cDNA,其长度为646 bp,包括一个258 bp开放阅读框;编码的GnRH前体为86个氨基酸残基,由一个信号肽、GnRH十肽和一个由蛋白水解位点(Gly-Lys-Arg)连接的促性腺激素释放激素相关肽(GAP)组成;其中信号肽和相关肽的长度分别为24和49个氨基酸。该cDNA编码的GnRH的前体氨基酸序列与其他物种的GnRH前体基本一致。表明物种间GnRHcDNA的蛋白编码区高度保守,而非编码区的保守性程度很低。进化分析发现,斑马鱼与鲤、鲫、拟鲤、黑头软口鲦等淡水鲤科鱼类的同源性较高。
The total RNA was extracted from brain of zebra fish. The GnRH cDNA was amplified by RT-PCR method using isolated total RNA as template. The GnRH cDNA is 646 bp in length that contain an open reading fram （ORF） of 258 bp, which encode the 86 amino acids residues that including a signal peptide （SP）, a GnRH decapeptide and a GnRH-associated peptide （GAP） which is linked by the processing site Gly- Lys-Arg sequence. The cGnRH cDNA encode SP of 24 amino acids residues and GAP of 49 amino acid residues. The sequence structure of amino acids precursor encoded by GnRH cDNA is basically identical with all others species reported to date. The result showed that the coding region of GnRH cDNA is highly conserved and the untranslated regions are markedly divergent in nucleotide sequence in the listed species. Phyligenetic tree indicated that Danio rerio was highly homologous with fresh water fish of Cyprinidae such as Cyprinus carpio , Carassius auratus , Rutilus rutilus , Pimephales promelas.
出处
《上海海洋大学学报》
CAS
CSCD
北大核心
2009年第3期263-268,共6页
Journal of Shanghai Ocean University
基金
青海省高校日元贷款人才培养项目(L/ANO.CO3-P189)
关键词
斑马鱼
促性腺激素释放激素
CDNA克隆
序列分析
Danio rerio
gonadotropin-releasing hormone （GnRH）
cDNA cloning
sequence analysis
作者简介
谢保胜（1965-），男，河南孟州人，硕士，副教授，主要从事细胞生物学与分子生物学方面的研究。E-mail：xbshjch@yahoo．com．cn
