黄牛白细胞介素-2基因的克隆与序列分析

CLONING AND SEQUENCE ANALYSIS ON IL-2 GENE IN CATTLE

为了克隆黄牛IL-2基因,采用RT-PCR技术,用ConA刺激12 h的黄牛颈部外周血淋巴细胞为材料,从总RNA中扩增出黄牛IL-2基因,琼脂糖凝胶电泳技术显示扩增片段约为500 bp,分离纯化,将其克隆到PDM18-T载体,重组质粒经PCR鉴定、双酶切鉴定以及核苷酸测序。结果显示,克隆的黄牛IL-2基因大小468 bp,与GenBank中报道的水牛、牛、绵羊、猫、马、鸡、犬、人、山羊、鼠、猪IL-2基因比较,同源性分别为98.9%;98.5%;97.0%;82.4%;79.9%;5.3%;78.2%;79.7%;95.9%;65.2%;84.7%。

In order to clone cattle interleukine-2 gene,according to GenBank in the bovine Interleukine-2（IL-2） cDNA sequences,a size of 500 bp cDNA fragment was amplified by RT-PCR from the total RNA extracted by ConA stimulated 12-hours cattle peripheral blood lymphocytes,and then constructed pMD-18T-IL-2 recombinant vector,following identification of PCR and double digestion of EcoR-1 and BamHI.Sequence analysis indicated that cattle IL-2 gene catained an open reading frames（ORF） of 468 bp.Compared the nucleotide with bovine,buffalo,sheep,cat,horse,chicken,canin,human,goat,mice and pig,the homoloaies were 98.9%,98.5%,97.0%,82.4%,79.9%,5.3%,78.2%,79.7%,95.9%,65.2% and 84.7%,respectively.