论文部分内容阅读
γ-氨基丁酸受体(GABAR)主要存在于脊椎动物和非脊椎动物的中枢神经系统(CNS),是氟虫腈、阿维菌素、硫丹和林丹等杀虫剂的作用靶标。为了阐明GABAR拮抗剂类农药影响鱼类安全性的分子作用机理,并研究其与农药分子的亲和作用,运用生物信息学方法,在NCBI基因组数据库中找到已经公布的同源的GABAARβ3亚基基因序列,通过多序列同源比对寻找高度保守区,再根据高度保守区设计特异性引物,通过RT PCR和RACE PCR技术,成功地克隆了鲫鱼GABAR A型β3亚基基因的全长cDNA序列(GenBank登录号:KC964110),该基因长2 767 bp,开放阅读框(ORF)为1 506 bp,编码502个氨基酸,与已知的其他物种β3亚基具有高度的保守性。应用qRT-PCR扩增,检测到该基因在鲫鱼不同组织器官的差异性表达。
γ-aminobutyric acid receptor (GABAR) is mainly found in the central nervous system (CNS) of vertebrates and invertebrates and is the target of insecticide such as fipronil, abamectin, endosulfan and lindane. In order to elucidate the molecular mechanism of GABAR antagonists affecting fish safety and to study their affinity with pesticide molecules, bioinformatics methods were used to find out the homologous GABAARβ3 subunit genes in the NCBI genome database Sequence, searching for highly conserved regions by multiple sequence homology alignment, and designing specific primers according to highly conserved regions. The full-length cDNA sequence of the GABAR A type β3 subunit gene was successfully cloned by RT PCR and RACE PCR GenBank accession number: KC964110). This gene was 2 767 bp in length with an open reading frame (ORF) of 1 506 bp encoding a polypeptide of 502 amino acids that was highly conserved from other known β3 subunits. QRT-PCR amplification was used to detect the differential expression of this gene in different tissues and organs of Carassius auratus.