CHARACTERIZATION OF THE CHICKEN PREPROGONADOTROPHIN-RELEASING HORMONE-I GENE
CHARACTERIZATION OF THE CHICKEN PREPROGONADOTROPHIN-RELEASING HORMONE-I GENE
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DOI：
10.1677/jme.0.0110019
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发表时间：
1993-08-01
影响因子：
3.5
通讯作者：
SANG, HM
中科院分区：
文献类型：
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作者：
DUNN, IC;CHEN, Y;SANG, HM
以鸡下丘脑总RNA为模板，采用逆转录-聚合酶链反应（RT-PCR）技术，克隆了鸡促性腺激素释放激素（GnRH）-I的部分cDNA。用于扩增的引物基于哺乳动物GnRH基因的核苷酸序列。将这些扩增的克隆用于筛选基因组文库，从该文库中分离出一系列重叠克隆。对含有所有外显子和3.0kb 5 ′上游区的6.3kb EcoRI片段进行测序。该基因的外显子-内含子结构被发现是一个类似的配置的哺乳动物和硬骨鱼类GnRH基因分析到目前为止。预测的prepropeptide的各个结构域与哺乳动物GnRH prepropeptide的结构域相似，包括23个氨基酸的信号肽、十肽激素和Gly-Lys-Arg切割位点，随后是56个氨基酸的GnRH相关肽。编码十肽激素的核苷酸序列翻译成鸡GnRH-I的氨基酸序列。前原肽与哺乳动物前原肽具有约50%的同一性，与硬骨鱼前原肽具有25%的同一性。
Partial cDNA clones for chicken gonadotrophin-releasing hormone (GnRH)-I were isolated by reverse transcription-polymerase chain reaction using total RNA from the hypothalami of domestic chickens. Primers for amplification were based on the nucleotide sequence of the mammalian GnRH genes. These amplified clones were used to screen a genomic library from which a series of overlapping clones was isolated. A 6.3 kb EcoRI fragment containing all the exons and 3.0 kb of the 5' upstream region was sequenced. The exon-intron structure of the gene was found to be of a similar configuration to those of the mammalian and osteichthyes GnRH genes analysed so far. Individual domains of the predicted prepropeptide are similar to those of mammalian GnRH prepropeptides, comprising a 23 amino acid signal peptide, the decapeptide hormone and a Gly-Lys-Arg cleavage site, followed by a 56 amino acid GnRH-associated peptide. The nucleotide sequence coding for the decapeptide hormone translates into the amino sequence for chicken GnRH-I. The prepropeptide has approximately 50% identity with mammalian prepropeptides and 25% identity with the teleost prepropeptides.