贵州黑山羊IL-2基因的克隆及序列分析

根据Gen Bank中山羊IL-2基因序列(登录号:NM_001287567)设计一对特异性引物,采用RT-PCR技术从贵州黑山羊外周血淋巴细胞中扩增IL-2基因,并将其克隆到p MD19-T载体上,构建IL-2基因克隆质粒,通过质粒PCR、双酶切和序列测定进行鉴定,对其核苷酸、氨基酸进行了比对分析并绘制系统进化树。结果表明,贵州黑山羊IL-2基因大小为468 bp,编码155个氨基酸。该基因与山羊、绵羊、藏羚羊、蓝牛羚、林麝、猪、牛、猫、大熊猫、犬、人、鼠、鸡等动物的核苷酸一致性在39.6%~100%,氨基酸一致性在26.7%~100%,遗传进化树表明,贵州黑山羊IL-2基因与鸡亲缘关系最远。

Cloning and sequence analysis of IL-2 gene of Guizhou black goat

Based on the published nucleotide sequence of caprine IL-2 gene( Accession No. NM_001287567) , a pair of primers IL-2 gene sequence were designed and synthesized. IL-2 gene was amplified from peripheral blood lymphocyte of Guizhou black goat by RT-PCR and cloned into pMD19-T vector to construct plasimid IL-2 gene. The nucleotide and a-mino acid were compared and analyzed through plasimid PCR, double enzyme digestion, sequence identification and con-struction of phylogeneric trees. The results showed that the IL-2 gene of Guizhou black goat was 468bp, encoding 155 a-mino acids. The IL-2 gene of Guizhou black goat was compared with the IL-2 of other goats: sheep, Tibetan antelope, nilgai, forest musk deer, pigs, cattle, cats, pandas, dogs, humans, mice, and chickens. The nucleotide consistency of Guizhou black goat with other animals is between 39. 6%~100% and the amino acid consistency is between 26. 7%~100%. Phylogenetic trees indicated that the IL-2 gene of Guizhou black goat is furthest away from chicken.