绵羊甘露聚糖结合凝集素基因(MBL)全长DNA序列克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Mannan-binding Lectin Gene(MBL) Full-length DNA Sequence in Sheep(Ovis aries)

作　　者：赵宗胜[1] 赵凤[1] 张红梅[2] 贾斌[1] 余鹏[3] 张慧[1] 班谦[1] 王建梅[1] 王小义[3] 

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]石河子大学医学院第一附属医院,石河子832002 [3]石河子大学生命科学学院,石河子832003

出　　处：《农业生物技术学报》2012年第12期1414-1423,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.30960247);科技部国际科技合作项目(No.2007DFB30420)

摘　　要：绵羊甘露聚糖结合凝集素(MBL)是肌体天然免疫系统的重要组成部分。根据人(Homo sapiens)和牛(Bos taurus)的MBL基因序列的同源保守区域,分段设计9对特异性引物,以中国美利奴羊(Ovis aries)血液基因组DNA为模板,经扩增、测序、拼接,首次克隆到长为4462bp的绵羊MBL基因组DNA序列,包括了该基因的启动子区、4个外显子和3个内含子,编码249个氨基酸;BLAST分析其与牛MBL-C编码的氨基酸同源性最高为96.39%,判定其为MBL-C型基因,并将该序列提交至GenBank(Accession No.FJ977629);通过Expasy网站预测其蛋白结构功能特征,1～19氨基酸为绵羊MBL基因信号肽区域,外显子1编码N-端富含胱氨酸区和8个Gly-X-Y重复序列的胶原样区,外显子2含有12个Gly-X-Y的胶原样区,外显子4编码的碳水化合物识别域(CRD)具有C型凝集素CRD的共同特征。本研究为国内首次报道绵羊MBL基因比较完整基因组DNA序列,为深入研究MBL的结构与功能以及MBL缺损的分子机制提供基础数据。Sheep mannose-binding lectin （MBL） is an important part of the body＇s natural immune system. Nine pairs of primers were designed according to human （Homo spiens） and bovine （Bos taurus） MBL gene sequence, combined with PCR technique to amplify the MBL gene of sheep （Ovis aries）. Cloning, sequencing and splicing to analyze bioinformations with the DNAMAN software and online tools. The results showed that sequence of DNA of the sheep MBL gene was 4 462 bp containing promoter, four exons, three introns, which encoded 249 animo acids, submitted it to GenBank （Accession No. FJ977629）. Its amino acid sequence had an overall similarity with a comparable region of bovine MBL-C gene （96.39%）, phylogenetic tree of amino acid analysis indicated that the sheep MBL gene was MBL-C. Expasy were employed to predict specific structureand function of sheep MBL gene, 1-19 amino acids were signal peptide region, exon 1 encoded N-terminus cysteine-rich domain and 8 Gly-X-Y repetitive sequences of collagen-like domain, and exon 4 encoded carbohydrate recognition domain （CRD） which had common features of C-type lectin. This study is the first report about complete genomic DNA sequence data of sheep MBL gene, and it provides basal data for further study on the structure, function and defects in the molecular mechanism of MBL.

关 键 词：绵羊 甘露聚糖结合凝集素(MBL) 序列分析 

分 类 号：Q98[生物学—人类学]