细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus

作　　者：刘田莉 孟庆玲[1] 乔军[1] 陈诚[1] 马玉[1] 胡政香 才学鹏[2] 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]中国农业科学院兰州兽医研究所,甘肃兰州730046

出　　处：《华北农学报》2015年第2期41-46,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家公益性(农业)行业专项(201303037);国家国际科技合作项目(2014DFR31310);新疆自治区研究生科研创新项目(No.XJGRI2014059)

摘　　要：为了研究绵羊细粒棘球蚴重要抗原基因Tetraspanin 1-TSP1(TSP1)和Tetraspanin 1-TSP6(TSP6)的功能,对Gen Bank中Eg基因组数据库检索,获得TSP1和TSP6的c DNA序列并设计特异性引物。以Eg头节为总RNA模板,进行RT-PCR,将PCR产物克隆到p MD19-T载体后测序并进行生物信息学分析。TSP1 c DNA全长792个核苷酸,编码263个氨基酸,该多肽含有3个潜在的N端糖基化位点,2个N端酰基化位点,与已登录的标准株TSP1基因序列(EG_11043)同源性为98.99%,其推导的氨基酸序列同源性为98.48%;TSP6 c DNA全长666个核苷酸,编码221个氨基酸,该多肽含有5个N端酰基化位点,1个酪氨酸激酶磷酸化位点,与已登录的标准株TSP6基因序列(EG_00715)同源性为98.18%,其推导的氨基酸序列同源性为85.07%。运用分子生物学软件对Eg TSP1和TSP6基因进行生物信息学分析,预测已知序列的蛋白质抗原的结构和表位,为疫苗的研发奠定了前期基础。In order to study the function of two important antigen genes tetraspanin 1-TSP1（ TSP1 ）and tet-raspanin 1-TSP6（TSP6）,primers derived from Echinococcus granulosus genome database in GenBank were designed and the open reading frame（ ORF）sequences of TSP1 and TSP6 were cloned by RT-PCR from hydatid protoscolex. Then they were cloned into pMD19-T vector for bioinformatics analysis. The results indicated that the TSP1 cDNA contains 792 nucleotides. The deduced protein consisted of 263 amino acids and has three N-glycosylation sites,two N-acylation sites. The gene sequence showed about 98. 99% identity with the TSP1（EG 11043）reported and the induced amino acid sequence showed about 98. 48% identity. The TSP6 cDNA contains 666 nucleotides. The de-duced protein consisted of 221 amino acids and has five N-acylation sites,one Tyrosine kinase phosphorylation sites. The gene sequence showed about 98. 18% identity with the TSP6（EG 00715）reported and the induced ami-no acid sequence showed about 85. 07% identity. The study carried out bioinformatics analysis of the TSP1 and TSP6 gene of Eg by molecular biology software to predict the structure and epitope of protein antigens known and laid a good foundation for the preparation of developing a vaccine.

关 键 词：细粒棘球蚴 抗原基因 基因克隆 生物信息学分析 

分 类 号：Q78[生物学—分子生物学]