为了确定γ-氨基丁酸B受体(gamma—aminobutyric acid B receptor,GABABR)基因在异育银鲫(Carassius auratus gibelio)不同组织中的表达,本实验分别对异育银鲫不同组织中GABABR1基因进行RT—PCR扩增,并进行了克隆和测序,在与GenBank基因库中已知GABA。R1序列进行同源性比对的基础上采用邻接法构建系统发育树,并进一步分析其在异育银鲫不同组织内的表达水平。经克隆获得异育银鲫GABABR1基因CDS区序列383bp,编码127个氨基酸。荧光定量PCR结果显示,GABABR1基因在异育银鲫脑、肝、肾、心、肠、鳔、鳃、肌、尾鳍、脾、卵巢、精巢组织中均有表达,且在不同组织中的表达水平由高到低依次是:脑〉尾鳍〉精巢〉心、肠、鳔〉卵巢、脾、鳃、肌〉肝、肾。本研究证实了GABABR1基因在异育银鲫各组织 表达的广泛性,且有明显的组织特异性。
To determine the expression of gamma-aminobutyric acid B receptor( GABABR) gene in different tissues of Carassius auratus gibelio, GABABR1 gene in different tissues was amplified by RT-PCR, cloned and sequenced, its phylogenetic tree was constructed using the neighbor-joining method on the basis of homology comparison with GABA~R1 amino acid sequences in the Gen Bank database, and additionally, the level of its expression in different tissues was further analyzed. The cDNA was 383 bp in length which encoded 127 amino acid residues. The real time PCR results showed that the GABABR1 gene was distributed in brain, liver, kidney, heart, intestine, muscle, bladder, gill, muscle, fin, spleen, ovary, and testis. The GABABR1 gene was expressed in different tissues, from high level to low level as follows: brain 〉 caudal fin 〉 testis 〉 heart, intestine, bladder ovary, spleen, gill, muscle 〉 liver, kidneys. The study confirmed the wide expression of GABABR1 gene in the tissues of C. auratus gibebio, with the highest expression in the brain tissue.