CAJ | 학술논문

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GABAAR β3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2767 bp，开放阅读框( ORF)为1506 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-안기정산수체( GABAR)주요존재우척추동물화비척추동물적중추신경계통( CNS)，시불충정、아유균소、류단화림단등살충제적작용파표。위료천명GABAR길항제류농약영향어류안전성적분자작용궤리，병연구기여농약분자적친화작용，운용생물신식학방법，재NCBI기인조수거고중조도이경공포적동원적GABAAR β3아기기인서렬，통과다서렬동원비대심조고도보수구，재근거고도보수구설계특이성인물，통과 RT PCR 화 RACE PCR 기술，성공지극륭료즉어GABAR A형β3아기기인적전장cDNA 서렬(GenBank 등록호： KC964110)，해기인장2767 bp，개방열독광( ORF)위1506 bp，편마502개안기산，여이지적기타물충β3아기구유고도적보수성。응용qRT-PCR 확증，검측도해기인재즉어불동조직기관적차이성표체。
γ-aminobutyric acid receptor ( GABAR ) mainly exist in the central nervous system of vertebrates and invertebrates ( CNS ) , it is the important targets of pesticides such as fipronil, abamectin, endosulfan and lindane. In order to clarify the molecular mechanism of GABA antagonists pesticides effects on the fishes, and the affinity with pesticide molecules, bioinformatic methods were used to search GABAA R β3 gene sequences that had released in the NCBI genome databases and did homologous alignment. Degenerate primers were designed in the highly conservative district. The full length cDNA sequence of GABAAR β3 of Carassius auratus was cloned successfully by RT-PCR and RACE PCR technology. The gene is 2 767 bp ( GenBank ID: KC964110 ) , with an open reading frame ( ORF) of 1 506 bp, encoding 502 amino acids. Multiple sequence alignment indicated that it was highly conserved with other known β3 subunits. QRT-PCR amplification was used to detect the activity of transcription of the gene in different tissues and organs of C. crassius. The results indicated that it was differentially expressed in organs of Carassius auratus.