鲤鱼肿瘤坏死因子α1基因cDNA的克隆及序列分析  

Cloning,Sequence Characterization Analysis of the Tumor Necrosis Factor alpha-1(TNFα1) Full-length cDNA from Carp

作　　者：赵晓[1] 何江帅[1] 冯祥汝[1] 陈义龙[1] 王彬[2] 李伟[3] 张俊辉[1] 王文东[1] 杨振国[2] 卢强[1] 

机构地区：[1]吉林大学人兽共患病研究所,吉林长春130062 [2]吉林大学畜牧兽医学院,吉林长春130062 [3]黑龙江生物科技职业学院,黑龙江哈尔滨150025

出　　处：《中国畜牧兽医》2011年第7期114-118,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30972277)

摘　　要：试验以鲤鱼外周血白细胞肿瘤坏死因子α1(tumor necrosis factor alpha 1,TNFα1)EST序列为基础,经地高辛标记作为探针对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行核酸杂交筛选,从0.9×104个重组噬菌体中,经过2轮筛选获得3个阳性克隆。序列分析结果显示,该序列包含128 bp的5′非编码区(5-′UTR),423 bp的3′非编码区(3-′UTR),开放阅读框ORF长768 bp,共编码255个氨基酸,在其3′非编码区存在几个ATTTA不稳定基序。预测蛋白等电点为8.20,分子质量大小为28.1 ku。序列同源性比较结果表明,所获序列与GenBank上登录的鲤鱼TNFα基因的同源性达94%。蛋白质的序列和结构分析结果发现,其具有TNF家族的典型序列特征、1个跨膜区结构和1个假定的产生成熟肽的裂解位点。The cDNA library of peripheral blood leucocytes which were separated from carp（Cyprinus carpio L.） and stimulated with mitogen was screened by a probe labelled with DIG.The tumor necrosis factor alpha（TNFα1） EST sequence was picked out from the cDNA library of peripheral blood leucocytes that was constructed.After two rounds of screening from 0.9×104 reeombinate phages,the positive clone was obtained.Sequence analysis indicated that it had a 128 bp 5′-UTR and a 423 bp 3′-UTR which included the instable motifs of ATTTA.The open reading frame（ORF）of this gene was 768 bp which putatively coded 255 amino acids with the MW of 28.1 ku,and PI of 8.20.Its homology with TNF from GenBank was up to 94%.Analysis of protein sequence and structure,within the predicted protein sequence we identified a TNF family signature,a predicted transmembrane domain and a putative cleavage site to produce the mature peptide.

关 键 词：鲤鱼 肿瘤坏死因子α1(TNFα1) 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]