鲤鱼肿瘤坏死因子受体相关因子6全长cDNA克隆及序列分析  

Clonging and Sequence Analysis of Tumor Necrosis Factor Receptor-associated Factor 6 Full-length cDNA from Common Carp

作　　者：贾生美[1] 孙真[1] 冯祥汝[1] 陈义龙[1] 沈雪飞[1] 翟新新[1] 张俊辉[1] 杨振国[1] 王文东[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

出　　处：《中国畜牧兽医》2013年第10期42-46,共5页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(30972277);吉林大学基本科研业务费项目(200903250)

摘　　要：本试验以鲤鱼外周血白细胞肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factor 6,TRAF6)EST序列为基础,经地高辛标记后作为探针,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,从重组噬菌体中经过两轮筛选获得阳性克隆。序列分析结果显示,该序列包含有5′-非编码区(5′-UTR)25bp;3′-非编码区(3′-UTR)535bp,存在2个mRNA不稳定基序ATTTA;开放阅读框ORF长1632bp,编码543个氨基酸。预测蛋白质等电点为5.88,分子质量大小为61.773ku。序列同源性比对结果表明,所获得的序列与GenBank上登录的鲤鱼TRAF6a基因同源性达99%。蛋白质序列分析结果发现,其具有TRAF家族的典型序列特征。The common carp tumor necrosis factor receptor-associated factor 6（TRAF 6）EST sequence was picked out from the cDNA library of peripheral blood leukocytes that was constructed.The cDNA library were separated from carp and stimulated with mitogen was screened by aprobe labeled with DIG.The full-length TRAF6 cDNA was cloned from recombinant phages.Sequence analysis indicated that it encompasses with a 25bp 5′-UTR and a 535bp 3′-UTR,the open reading frame（ORF）of which was 1632bp putatively coding 543 amino acids, and there were two motifs for mRNA instability ATTTA in the 3′-untranslated region.The predicted theoretical isoelectric point and molecular weight were 5.88 and 61.773ku,respectively.Its nucleotide homology with carp TRAF6 from GenBank was up to 99%.The protein sequence analysis showed that it shared typical sequence features of the TRAF family.

关 键 词：鲤鱼 肿瘤坏死因子受体相关因子6 克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]