细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

摘要

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.%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GenBank中 Eg基因组数据库检索，获得 TSP1和 TSP6的 cDNA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 pMD19-T载体后测序并进行生物信息学分析。TSP1 cDNA 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 cDNA 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。