细粒棘球蚴<em>TSP1</em>和<em>TSP6</em>基因的克隆及生物信息学分析

doi:10.7668/hbnxb.2015.02.008

华北农学报 ›› 2015, Vol. 30 ›› Issue (2): 41-46. doi: 10.7668/hbnxb.2015.02.008

所属专题： 生物技术；

细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

刘田莉¹, 孟庆玲¹, 乔军¹, 陈诚¹, 马玉¹, 胡政香¹, 才学鹏², 陈创夫¹

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

收稿日期:2015-02-10 出版日期:2015-04-28
作者简介:刘田莉(1992-),女,河南商丘人,在读硕士,主要从事动物寄生虫研究。
基金资助:
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)

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

LIU Tian-li¹, MENG Qing-ling¹, QIAO Jun¹, CHEN Cheng¹, MA Yu¹, HU Zheng-xiang¹, CAI Xue-peng², CHEN Chuang-fu¹

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

Received:2015-02-10 Published:2015-04-28

摘要/Abstract

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。

关键词: 细粒棘球蚴, 抗原基因, 基因克隆, 生物信息学分析

Abstract: In order to study the function of two important antigen genes tetraspanin 1-TSP1(TSP1) and tetraspanin 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 deduced 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 amino 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.

Key words: Cystic echinococcosis, Antigen gene, Gene cloning, Bioinformatics analysis

中图分类号:

刘田莉, 孟庆玲, 乔军, 陈诚, 马玉, 胡政香, 才学鹏, 陈创夫. 细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析[J]. 华北农学报, 2015, 30(2): 41-46. doi: 10.7668/hbnxb.2015.02.008.

LIU Tian-li, MENG Qing-ling, QIAO Jun, CHEN Cheng, MA Yu, HU Zheng-xiang, CAI Xue-peng, CHEN Chuang-fu. Cloning and Bioinformatics Analysis of TSP1 and TSP6 Gene of Echinococcus granulosus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(2): 41-46. doi: 10.7668/hbnxb.2015.02.008.

http://www.hbnxb.net/CN/Y2015/V30/I2/41

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细粒棘球蚴TSP1和TSP6基因的克隆及生物信息学分析

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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

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