Cloning And Analysis Of GABA Subunit Subfamily Gene Of Carassius Auratus

y-aminobutyric acid receptor (GABAR) mainly exist in the central nervous system of vertebrates and invertebrates (CNS)，GAB A receptors are the target of pesticide such as ifpronil. abamectin, endosulfan and lindane, but also thepharmaceutical target of epilepsy, alcoholism, schizophrenia and Parkinson’s disease, so GAB A receptors are highly valued. Experimental studies have shown that avermectin, fipronil and other pesticides are highly toxic to fish, Our studies were aimed at cloning and purifying the GABA receptor subunit of fish，and studying the amino acid sequence and its three-dimensional structure.Bioinformatic methods were used to search GABAaR p3gene sequences thathave released in the NCBI genome databases and did homologous alignment，Thedegenerate primers were designed in the highly conservative district，The full lengthcDNA sequence of GABAaR p3of Carassius auratus was cloned successfully by RTPCR and RACE PCR technolog. The gene is2765bp (GenBank ID: KC964110). withan open reading frame (ORF) of1506bp, encoding502amino acids. Multiplesequence alignment has been done and found it was highly conserved with otherknown^subunits. QRT-PCR amplification was used to detect the activity oftranscription of the gene in different tissues and organs of Crassius crassius，theresults indicated that it was differentially expressed in organs of Carassius auratus.We used the avermectin-sensitive glutamate-gated chloride channel a-subunit of thereceptor protein (PDB1D:3RHW) as a template, simulated homologous pentamerstructure of p3subunit, optimized the structure model and calculated the mode ofaction and the binding capacity. Studying the effect between the fish GABAaRsubunits and pesticide molecular It Seeks to clarify the molecular mechanism ofGABA antagonists pesticides effects on the ifshes. Laid a certain foundation for thefurther research of the effects between GABAaR p3subunit and the pesticides and thecloning of GABA receptors.