七鳃鳗(Lampetra japonica)白细胞cDNA文库构建及生物信息学分析  

The construction of Leukocyte cDNA library and bioinformatics analysis of Lampetra japonica

作　　者：张宁[1,2] 刘欣[2,3] 孙晶[2] 刘岑洁[2] 陈操[2] 李竞[2] 芦静[2] 刘慧玲[2] 李庆伟[2] 

机构地区：[1]辽宁师范大学科研处,辽宁大连116029 [2]辽宁师范大学海洋生物功能基因及蛋白质组学研究所,辽宁大连116029 [3]辽宁师范大学城市与环境学院,辽宁大连116029

出　　处：《辽宁师范大学学报（自然科学版）》2010年第2期237-243,共7页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家重点基础研究发展计划项目(2007CB815802);国家高技术研究发展计划项目(2007AA09Z400);国家自然科学基金项目(30970342);教育部留学回国人员基金项目;辽宁省高校创新团队支持计划项目(2006R32;2007T089;2008T103)

摘　　要：以复合免疫日本七鳃鳗(Lampetra japonica)外周血中分离的单个核白细胞为材料,构建库容量为4.5×106pfu.mL-1的cDNA文库,随机挑选单克隆菌落测序共得到10 500条有效ESTs(expressed sequence tags)序列.经PHRAP软件对序列拼接组装后得到3 382个转录本,其中有1 080个重叠群和2 302个单一序列,有59.08%的转本录在NCBI数据库中搜索到同源序列.运用Blast2GO软件进行Gene Ontology注释,分别有27.47%、25.20%和10.01%的转录本参与细胞生理、代谢和生物调节等生物学途径;分别有48.43%、22.10%和10.69%的转录本起到结合、催化和结构分子活性等分子功能.统计数据显示,与细胞自身基因转录、蛋白质合成及转运等生命过程密切相关的基因如真核转录延长因子、反转录酶、核糖体蛋白、腺苷酸转运体、细胞色素C氧化酶和热休克蛋白等在血液白细胞群中大量表达.另外,与固有免疫密切相关的白细胞衍生趋化因子、抵抗素、颗粒体蛋白等表达量相对较高,说明固有免疫系统在七鳃鳗感染初期的免疫防卫方面起着主要作用.A cDNA library with 4.5×106 pfu·mL-1 capability was constructed from the mononuclear leukocytes extracted from peripheral blood of Lampetra japonica which were immunized with antigen complexs,and more than 10 500 randomly selected clones were sequenced.These sequences were contiged by PHRAP software and 3 382 transcripts are obtained.There are 1 080 consensus sequences and 2 302 unique transcripts,59.08% transcripts shared similarity to sequences published in NCBI databases.The functional annotations to assemble ESTs with the Blast2GO software showed that 27.47%、25.20% and 10.01% transcripts involved in cell physiology,metabolism,physiological modulation;and 48.43%、22.10% and 10.69% transcripts play important roles in binding or catalyzing functional polymers respectively.The data indicate the excessive expression of genes,including transcription elongation factor,reverse transcriptase,ribosome protein gene,adenine nucleotide translocator,cytoehrome oxidase and heat shock protein,are closely related to cell gene transcription activities and protein synthesis and transport.In addition,the researches demonstrate the highly expressed genes of chemokine,resistin and granulin are related to innate immunity.The findings thus suggest that innate immunity system plays a major role in early infection of the lamprey immune system.

关 键 词：日本七鳃鳗 类淋巴细胞 EST 分子标识 发育分化 

分 类 号：Q811.4[生物学—生物工程] Q959.392