›› 2015, Vol. 42 ›› Issue (10): 2631-2635.doi: 10.16431/j.cnki.1671-7236.2015.10.018

• 生物技术 • 上一篇    下一篇

副猪嗜血杆菌丝氨酸蛋白酶基因的克隆及生物信息学分析

游锦洲1, 张瑞振1, 丘建龙2, 谢敏1, 胡玲英1,2   

  1. 1. 龙岩出入境检验检疫局, 龙岩 364000;
    2. 福建农林大学动物科学学院, 福州 350002
  • 收稿日期:2015-03-30 出版日期:2015-10-20 发布日期:2015-10-23
  • 通讯作者: 胡玲英 E-mail:lingyingh@yeah.net
  • 作者简介:游锦洲(1989-),男,福建永定人,学士,研究方向:预防兽医学,E-mail:youjinzhou06@126.com
  • 基金资助:
    福建省科技厅省属高校项目(JK2011052)

Cloning and Bioinformatics Analysis of espP Gene of Haemophilus parasuis

YOU Jin-zhou1, ZHANG Rui-zhen1, QIU Jian-long2, XIE Min1, HU Ling-ying1,2   

  1. 1. Longyan Entry-exit Inspection & Quarantine Bureau, Longyan 364000, China;
    2. College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2015-03-30 Online:2015-10-20 Published:2015-10-23

摘要: 为研究副猪嗜血杆菌(Haemophilus parasuis,Hps)丝氨酸蛋白酶(espP)作为疫苗候选抗原的可能性,根据GenBank中Hps espP基因(登录号:HAPS_1381)设计引物,以Hps血清11型(H456)菌株的DNA为模板,PCR扩增目的基因并测序。将该基因核苷酸序列翻译成氨基酸序列后,对Hps espP蛋白的信号肽、亲水性区域和B细胞抗原表位进行预测,并预测了该蛋白C端的3D结构。结果显示,Hps espP基因序列全长2 343 bp,共可编码781个氨基酸,其中氨基酸序列的1—23位预测为信号肽。Hps espP蛋白含有12个亲水性区域,并预测含有12个B淋巴细胞抗原表位。构建Hps espP蛋白3D结构显示,该蛋白C端是由12个β折叠层围成的桶状结构。本试验为进一步研制新型副猪嗜血杆菌Hps espP蛋白多肽疫苗奠定基础。

关键词: 副猪嗜血杆菌; 丝氨酸蛋白酶; 克隆; 生物信息学

Abstract: In order to evaluate whether Haemophilus parasuis espP gene could use as a vaccine candidate antigen or not,the espP gene was amplified by PCR using the specific primers that designed according to corresponding sequence getting form GenBank (accession No.:HAPS_1381) and then sent to sequence.After translating the DNA sequence into amino acid sequence,the signal peptide,hydrophilic region,linear B cell epitopes and 3D structure of Hps espP protein were predicted using multiple bioinformatics analysis.The results showed that the full length of Hps espP gene was 2 343 bp,encoding 781 aa.The first 23 aa were predicted as signal peptide.There were 12 hydrophilic regions and 12 B cell epitopes in the Hps espP protein.The 3D structure of C-terminal of the protein was consisted of a monomeric β-barrel containing 12 β-strands and a central pore.The study laid the foundation for further development of molecular vaccines based on espP protein against Hps infection.

Key words: Haemophilus parasuis; espP; clone; bioinformatics

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