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作 者:胡贵川[1] 冯将[1] 鲜思美[1,2] 刘嫒[1] 李鹏飞[1] 肖拉拉 HU Gui-chuan FENG Jiang XIAN Si-mei LIU Ai XIAO La-la(Animal Science College of Guizhou University, Guiyang 550025, China Guizhou Insitute of Animal Disease, Guiyang 550025, China)
机构地区:[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病研究所,贵州贵阳550025
出 处:《山地农业生物学报》2016年第5期25-29,36,共6页Journal of Mountain Agriculture and Biology
基 金:贵州大学2014年大学生"SRT计划"项目(贵大SRT字[2014]124号);贵州省科技计划课题(黔科合LH字[2014]7667)
摘 要: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.
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