白来杭鸡IL-18和GM-CSF基因克隆与序列分析  

CLONING AND SEQUENCE ANALYSIS OF IL-18 AND GM-CSF GENE OF GALLUS GALLUS BREED WHITE LEGHORN

作　　者：于新友 赵蕾[1,2] 孙翠萍 沈志强[1,2] 

机构地区：[1]山东绿都生物科技有限公司,滨州256600 [2]山东省滨州畜牧兽医研究院,滨州256600

出　　处：《中国动物传染病学报》2011年第3期28-33,共6页Chinese Journal of Animal Infectious Diseases

基　　金：山东省自主创新成果转化重大专项计划(2008ZHZX1A1103)

摘　　要：根据GenBank中鸡白细胞介素18(inteleukin 18,IL-18)和粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)基因序列,克隆白来杭鸡IL-18和GM-CSF基因并进行序列分析。结果显示:白来杭鸡IL-18基因开放阅读框为597 bp,编码199个氨基酸,所获得的白来杭鸡IL-18基因与GenBank鸡IL-18的核苷酸同源性为99.0%～99.8%,氨基酸同源性为97.5%～99.5%。白来杭鸡GM-CSF基因开放阅读框为435 bp,编码145个氨基酸,所获得的白来杭鸡GM-CSF基因与GenBank鸡GM-CSF的核苷酸同源性为99.3%～100%,氨基酸同源性为99.3%-100%。IL-18与GM-CSF基因编码的蛋白具有亲水性,都有很强的抗原性。IL-18共有2个潜在的N-糖基化位点,可能存在4个蛋白激酶C磷酸化位点及3个酪蛋白激酶Ⅱ磷酸化位点。GM-CSF共有1个潜在的N-糖基化位点,可能存在1个蛋白激酶C磷酸化位点及4个酪蛋白激酶Ⅱ磷酸化位点。本研究结果为进一步研究鸡IL-18、GM-CSF基因表达、生物学活性和应用奠定了基础。Two pairs of primers were designed,based on the sequences of inteleukin 18（IL-18） and granulocyte-macrophage colony stimulating factor（GM-CSF） gene of Chicken on GenBank website.Total RNA of spleen lymphocyte of Gallus gallus breed White Leghorn was taken as template for RT-PCR.IL-18 and GM-CSF genes in Gallus gallus breed White Leghorn were cloned and sequenced.The sequencing results showed that the open reading frame（ORF） of IL-18 gene was 597 bp,coding 199 amino acids.The sequence analysis showed the homology of IL-18 gene between Gallus gallus breed White Leghorn and other chicken were 99.0%-99.8% at nucleotide level,and were 97.5%-99.5% at amino acid level.The ORF of GM-CSF gene was 435 bp,coding 145 amino acids.The sequence analysis showed the homology of GM-CSF gene between Gallus gallus breed White Leghorn and other chicken on GenBank website were 99.3%-100% at nucleotide level,and were 99.3%-100% at amino acid level.Bioinformatics analysis of this sequence showed that the protein of ChIL-18 and ChGM-CSF were hydrophilic and highly antigenic.IL-18 included two potential N-glycosylation site,four potential proteinkinase C phosphorylation site,and three potential caseinkinaseⅡ phosphorylation site.GM-CSF included one potential N-glycosylation site,one potential proteinkinase C phosphorylation site,and four potential caseinkinaseⅡ phosphorylation site.This study established the way for future study on biological function and application of chicken IL-18 and GM-CSF.

关 键 词：白细胞介素18 粒细胞-巨噬细胞集落刺激因子 序列分析 

分 类 号：S858.312.4[农业科学—临床兽医学] Q7[农业科学—兽医学]