Wang Cui, Liu Yi, Wang Huiying, Wu Huali, Gong Shaoming, He Daqian
National Poultry Engineering Research Centre, Institute of Animal Husbandry and Veterinary Science, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, People's Republic of China.
Mol Biol Rep. 2014 May;41(5):2929-36. doi: 10.1007/s11033-014-3149-y. Epub 2014 Jan 23.
Dopamine D1 receptor (DRD1) is one of the dopamine receptors with seven transmembrane domains that are coupled to the G protein. In the present study, we cloned the full coding region of DRD1 gene by the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends from the goose hypothalamus tissues. Results showed that the goose DRD1 cDNA (GenBank: KF156790) contained a 1,356 bp open reading frame encoding a protein 452 amino acid with a molecular weight of 50.52 kDa and a isoelectric point of 6.96. Bioinformatics analysis indicated that the deduced amino acid sequence was 71-98% identical to the DRD1 protein of other species, contained seven transmembrane domains and four N-glycosylation sites. A phylogenetic tree analysis revealed that the deduced goose DRD1 protein had a close genetic relationship and evolutional distance with that of duck, chicken, and zebra finch. The semi-quantitative RT-PCR analysis displayed goose DRD1 gene was widely expressed in all detected tissues, including heart, lung, liver, spleen, kidney, breast muscle, duodenum, sebum, pituitary, hypothalamus, ovary and oviduct. Eighteen single nucleotide polymorphisms were indentified in 3,169 bp length of this gene. For G90A mutation, the genotyping analysis of PCR-TspRI-RFLP showed the allele G was in dominance in all detected goose breeds, and the allele frequencies of this polymorphism were significantly different between Chinese goose breeds and foreign breeds (P<0.01). These findings will help us understand the functions of the DRD1 gene and the molecular breeding in geese.
多巴胺 D1 受体(DRD1)是一种具有七个跨膜结构域且与 G 蛋白偶联的多巴胺受体。在本研究中,我们通过逆转录聚合酶链反应(RT-PCR)和从鹅下丘脑组织中快速扩增 cDNA 末端,克隆了 DRD1 基因的完整编码区。结果表明,鹅 DRD1 cDNA(GenBank:KF156790)包含一个 1356 bp 的开放阅读框,编码一个由 452 个氨基酸组成的蛋白质,分子量为 50.52 kDa,等电点为 6.96。生物信息学分析表明,推导的氨基酸序列与其他物种的 DRD1 蛋白具有 71%-98%的同一性,包含七个跨膜结构域和四个 N-糖基化位点。系统发育树分析显示,推导的鹅 DRD1 蛋白与鸭、鸡和斑胸草雀的 DRD1 蛋白具有密切的遗传关系和进化距离。半定量 RT-PCR 分析显示,鹅 DRD1 基因在所有检测组织中广泛表达,包括心脏、肺、肝脏、脾脏、肾脏、胸肌、十二指肠、皮脂、垂体、下丘脑、卵巢和输卵管。在该基因 3169 bp 的长度中鉴定出 18 个单核苷酸多态性。对于 G90A 突变,PCR-TspRI-RFLP 的基因分型分析表明,等位基因 G 在所有检测的鹅品种中占优势,并且该多态性的等位基因频率在中国鹅品种和外国品种之间存在显著差异(P<0.01)。这些发现将有助于我们了解 DRD1 基因的功能以及鹅的分子育种。