水牛TLR2cDNA的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of TLR2 cDNA in Buffalo

作　　者：张晓茹[1,2] 匡文华[1,2] 成鹰[1] 杜丽[1] 张冬琳[1] 郝永昌[1] 雷明[1] 刘涛[1] 焦寒伟[1] 王凤阳[1] 祁超[2] 

机构地区：[1]海南大学农学院海南省热带动物繁育与疫病研究重点实验室(筹)海口市动物基因工程重点实验室,海南海口570228 [2]华中师范大学生命科学学院,湖北武汉430079

出　　处：《动物医学进展》2011年第11期35-41,共7页Progress In Veterinary Medicine

基　　金：国家转基因生物新品种培育重大专项基金(2009ZX08007-009B)

摘　　要：本研究通过RT-PCR分段扩增得到水牛Toll样受体2(TLR2)cDNA序列,并利用亚克隆重叠区进行全长序列拼接。将拼接产物与pMD20-T载体连接,重组质粒经PCR酶切鉴定后测序,并进行生物信息学分析。结果表明,克隆得到的序列全长2 455bp,含有一个大小为2 355bp的开放阅读框,共编码784个氨基酸,分子质量为90.2ku,理论等电点为6.92;TLR2ORF序列与GenBank上登载的水牛核苷酸序列同源性为99.53%,与黄牛、马、人、黑猩猩和狗的核苷酸序列同源性分别为97.88%、84.94%、83.12%、82.95%和82.74%,具有很高的相似性;该蛋白是一个跨膜蛋白,存在LRR、LRR-TYP、LRRCT及TIR结构域,与Toll样受体家族成员的共同结构相一致。水牛TLR2基因的成功克隆及序列分析和结构预测为进一步研究其生物学功能奠定了基础。In this report,Toll-like receptor 2（TLR2） cDNA of buffalo was amplified partly by RT-PCR.The amplified fragments were spliced into a full-length fragment through the overlapping areas.The splicing product was then cloned into the pMD20-T vector,the recombinant plasmid was confirmed by PCR and endonuclease digestion,then it was sequenced and bioinformatically analyzed.Results showed that TLR2 cDNA was 2 455 bp in length and contained a 2 355 bp ORF,encoding 784 amino acids,with the molecular weight of 90.2 ku,and the theoretical IP was 6.92.The nucleotide sequence of the cloned TLR2 ORF shared 99.53% homologies with that of buffalo published in GenBank.And the homology between the nucleotide sequence of the cloned TLR2 ORF and that of cattle,horse,human,chimpanzee and dog were 97.88%,84.94%,83.12%,82.95% and 82.74% respectively.These data suggested that TLR2 gene was relatively conserved among different species.TLR2 is a transmembrane protein,contains several domains like LRR,LRR-TYP,LRRCT and TIR,which are consistent with the common structure of Toll-like receptor family members.The successful clone,sequence analysis and structure prediction of TLR2 cDNA of buffalo lays a foundation for further research on the biological function of the gene.

关 键 词：TOLL样受体2 水牛 分子克隆 生物信息学分析 

分 类 号：S852.4[农业科学—基础兽医学] S858.23[农业科学—兽医学]