犬GM-CSF基因的克隆、序列分析及蛋白结构预测

Cloning and sequence analysis of Canis GM-CSF gene and prediction of protein structure

为了研究犬粒细胞/巨噬细胞集落刺激因子(GM-CSF)的作用,试验根据GenBank上发表的犬(Canis familiaris)GM-CSF cDNA基因序列设计1对引物,采用RT-PCR技术,以ConA刺激的犬外周血淋巴细胞为材料,从总RNA中扩增出犬GM-CSF基因。结果表明:扩增片段长为435 bp,编码144个氨基酸;生物软件分析结果表明,该序列与猕猴的亲源性更近,与已知人、马、牛、野猪、豹猫、鸡等序列的同源性较低。

In order to research the function of canine GM-CSF,based on the published nucleotide sequence of canine（Canis familiaris）GM-CSF gene,a pair of primers were designed and synthesized,the total RNA was isolated from ConA-stimulated canine peripheral blood T cells,then the CaGM-CSF gene was cloned by RT-PCR technique.The result showed that cloned GM-CSF cDNA was 435 bp in size and encoded a polypeptide of 144 amino acids.Compared CaGM-CSF with published on GenBank.The high identity between CaGM-CSF and Rhesus macaque was found,but low identity was found among canine GM-CSF,human GM-CSF,horse GM-CSF,buttle GM-CSF,pig GM-CSF,cat GM-CSF,chicken GM-CSF and so on.The cloned CaGM-CSF gene may construct a basis on further research on protein fuction.