合作猪TLR3及其剪接体的基因克隆与序列分析  

Cloning and sequence analysis of TLR3 and its spliceosomes in Hezuo pig

作　　者：管齐赛[1] 房永祥[1] 贾怀杰[1] 何小兵[1] 周涛[1] 景志忠[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点实验室甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《中国兽医科学》2012年第7期731-736,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30871884);国家高技术研究发展计划(863)项目(2011AA10A211)

摘　　要：为了研究猪Toll样受体3(pTLR3)基因的选择性剪切机制,应用反转录-聚合酶链反应扩增技术,从猪外周血淋巴细胞中克隆了pTLR3及其剪接体基因(pTLR3a,pTLR3b)。pTLR3基因序列分析表明,克隆的基因ORF为2 718bp,编码905个氨基酸;推导的氨基酸序列分析显示,在其胞外区具有LRRs结构域,胞内区含有TIR结构域,具有TLR家族的典型结构特征。剪接体分析显示,pTLR3a和pTLR3b基因缺失第3外显子,都没有可编码跨膜区、胞内区的部分。相似性分析显示,与牛、马、猫和人的相似性较高,与家兔、鼠的相似性次之。证实pTLR3可变剪接体的存在为进一步研究其功能奠定了基础。The objective of the present study was to investigate the alternative splicing of porcine Toll- like receptor 3（pTLR3） gene in Hezuo pig. The pTLR3 gene and spliceosomes（pTLR3a and pTLR3b） were cloned from porcine peripheral blood mononuclear cells by reverse transcription-polymerase chain re- action. Sequence analysis showed that the pTLR3 open reading frame was 2 718 bp in length encoded a de- duced protein with 905 amino acid residues. The sequence analysis of deduced amino acids indicated that it was a typical type I transmembrane protein with a LRRs ectodomain and a TIR cytoplasmic domain, the conserved structural features of TLRs. Due to the absence of extron 3, the spliceosomes of pTLR3a and pTLR3b could not encode the transmembrane and cytoplasmic domains. The similarity of nucleotide se- quence of pTLR3 with those of cattle, equine, cat and human was much higher than that of rabbit and mice. It was concluded that the alternative spliceosomes of pTLR3 do exist,and which lay foundations for further studies on their molecular functions.

关 键 词：猪TLR3 克隆 序列分析 可变剪接 

分 类 号：Q51[生物学—生物化学]