【摘要】 为深入研究鸡G蛋白α亚基(G-protein alpha subunit, GNAS)基因的转录剪接情况,采用5’和3’cDNA末端快速扩增(rapid-amplification cDNA ends, RACE)技术,对鸡GNAS基因进行克隆测序;并采用定量聚合酶链式反应(polymerase chain reaction, PCR)技术,检测鸡皮肤、胸肌、心、脑、肝、肺和腹脂等7种组织中GNAS基因剪接体的表达量。结果表明:鸡GNAS基因存在1 554和1 796 bp 2种转录剪接体,均含12个外显子,二者仅第1外显子的长度和位置不同,其余第2-12外显子相同。克隆子1(1 554 bp)编码417个氨基酸,克隆子2(1 796 bp)编码379个氨基酸。在NCBI数据库中进行蛋白比对发现,克隆子2的氨基酸序列与已知的人和小鼠的GNAS基因Gαs亚基同源相似度达93%,而克隆子1的氨基酸序列与XLαs亚基的同源性较高(相似度87%)。表达量检测表明,2种转录剪接体在7种组织中均有不同程度的表达,其中:在脑组织中的表达量最高,与其他组织间差异极显著(P<0.01),在皮肤组织中的表达量(P<0.01或P<0.05)次之;而在肺、胸肌、肝、心、腹脂等组织中的表达量较低。更多还原

【Abstract】 In order to research deeply the transcription and splicing situations of chicken G-protein alpha subunit gene(GNAS), we used 5’ and 3’ rapid-amplification cDNA end(RACE) technology to clone and sequence the chicken GNAS gene, and used quantitative polymerase chain reaction(PCR) to detect the expression level of GNAS spliceosome in seven tissues of chicken,such as skin, pectoral muscle, heart, brain, liver, lung,and abdominal fat. The results showed that the chicken GNAS gene had two transcriptional spliceosomes of 1 554 bp and 1 796 bp, respectively. Both of spliceosomes included 12 exons, and only the length and position of their first exon were different, and the second to twelfth exons were the same. Clone 1(1 554 bp) coded 417 amino acids, while clone 2(1 796 bp) coded 379 amino acids. Protein alignment in the NCBI database showed that the similarity between the 379 amino acid sequence of the clone 2 and Gαs subunits of the known human and mouse GNAS genes was 93%. The 417 amino acid sequence of the clone 1 was more similar with the XLαs subunit, and their similarity was 87%. The gene expression detection showed that these two transcriptional spliceosomes had different degrees of expression in the seven tissues: the highest expression in the brain(P<0.01), next in the skin(P<0.01 or P<0.05), and the lower expressions in the lung, pectoral muscle, heart and abdominal fat.更多还原