贵州黑山羊IL-2基因的克隆及序列分析

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

根据GenBank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pMD19-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。

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.