目的:进行美国王鸽脑组织中GABAA受体α1亚基、β2亚基和γ2亚基的基因序列的克隆与分析。方法:从美国王鸽大脑中提取总RNA,应用通用引物逆转录获得基因组cDNA。设计GABAA受体三种亚基的特异引物,通过RT-PCR分别扩增出GABAA受体α1亚基、β2亚基和γ2亚基的基因序列,并对三段基因序列进行克隆、质粒鉴定及分析。结果:成功地测出GABAA受体α1亚基、β2亚基和γ2亚基的基因序列,长度依次为899bp、597bp和564bp,并得出其与Gallusgallus参考序列的同源率分别为94.99%、94.64%和96.28%。结论:鸽子脑组织中GABAA受体α1亚基、β2亚基和γ2亚基基因序列与已知原红鸡相应基因序列同源性较高,但也显示出一定的种属差别。
Aim: To clone and analyse the coding cDNA sequence of α1、β2 and γ2subunit of GABAA receptor in American king Pigeon. Methods: Withdrew total RNA from the American king pigeon brain, reverse transcribing general primers to acquire a gene set cDNA.Designing specific primers of three subunit mRNA of the GABAA receptor,by RT-PCR respectively expanded the conservative gene of α1 subunit, β2 subunit and γ2 subunit of GABAA receptor, and carried on clone, plastid identification and the sequence measurese of three genes. Results: The experiment on sequence measurese has succeeded that sequence analysis indicated that lengths of the conservative gene of α1 subunit, β2 subunit and γ2 subunit of GABAA receptor was respectively 899 bp, 597 bp and 563 bp, homologoy on reference sequence was respectively 94.99%, 94.64% and 96.28%. Conelusien: Homology is high on the conservative gene of α1 suhunit, β2 subunit and γ2 subunit of GABAA receptor of brain tissue of pigeon and chicken but there is a discriminating characteristic in different kinds of animals.