猪(杜长大三元杂)白细胞介素-6基因的克隆及序列分析  

Gene cloning and sequence analysis of swine (crossbreeding pig) interleukin-6

作　　者：张桂红[1] 林海波[1] 江经纬[1] 詹国英[1] 廖明[1] 

机构地区：[1]华南农业大学兽医学院禽病室,广东广州510642

出　　处：《黑龙江畜牧兽医》2006年第7期18-20,共3页Heilongjiang Animal Science And veterinary Medicine

基　　金：广东省科技攻关项目(粤科计字【2003】292号);华南农业大学校长基金(K03005)

摘　　要：分离猪（杜长大三元杂）外周血单核细胞（PBMC）进行体外培乔，经诱导刑LPS或ConA的诱导培养48h后，提取细胞总RNA，应用RT-PCR扩增猪IL-基因，并克隆到pMDl8-T载体后测序。结果成功获得猪IL-6基因重组质粒pMDpIL-6，序列分析发现所获得的猪IL-6cDNA长度为716bp．开放阅读框为636bp，编码211个氨基酸，相对分子质量约24ku，等电点5．5。所获得的猪IL-6与GenBank上已报道的不同来源猪的IL-6有单个氨基酸存在差异；与人和其他动物IL-6氨基酸序列的同源性为78．1％-90．1％，同时反映出IL-6基因具有种的多样性，不同种动物的IL-6基因亲缘关系越近，进化关系也越近。The peripheral blood monocytes of swine（Cross-breeding Pig） were isolated and cultured in vitro Whereafter we extracted the total RNA from the cells and amplified the gene of interleukin-6 with reverse transcription polymerase chain react/on（RT- PCR）after the ceils were incubated with the proper inducer（LPS or ConA） for 48 hours. The RT -PCR product was cloned nto the pMD18-T vector,and the extracted plasmid was sequenced after then. The re-sult indicated that a recombinant plasmid（ denoted as pMDpIL -6）including the interleukin-6 gene was obtained,Sequence analysis indicated that the cDNA of the interleukin-6 gene was 716bp in length, where the open reading frame （ORF） was 636bp in length encoding 211 amino acids which was 24KD in molecular weight,and the isoelectric point of the interleukin-6protein was 5.5. The sequence of recombinant interleukin-6 was compared with those from the swine of different origins in Genbank, and it only differed in one amino acid. Then, the recombinant interleukin-6 was aligned with those from the different species in Genbank,and the homology of the sequence of amino acid residues ranged from 78. 1% to 90. 1%. Meanwhile,our result suggested that the inter-leukin-6 gene was of species diversity, and that different animals were closer on the phylogenetic tree while their interleukin-6 genes were closer in sequence distances.

关 键 词：猪 白细胞介素-6 基因克隆 序列分析 

分 类 号：S828[农业科学—畜牧学]