中国媒介生物学及控制杂志 ›› 2016, Vol. 27 ›› Issue (6): 533-538.DOI: 10.11853/j.issn.1003.8280.2016.06.002

• 论著 • 上一篇    下一篇

新发虫媒病毒:基因A型版纳病毒全基因组序列扩增引物

刘红1,2, 梁国栋2   

  1. 1. 山东理工大学生命科学学院生物技术系, 山东 淄博 255000;
    2. 中国疾病预防控制中心病毒病预防控制所病毒性脑炎室, 传染病预防控制国家重点实验室, 北京 102206
  • 收稿日期:2016-07-21 出版日期:2016-12-20 发布日期:2016-12-20
  • 通讯作者: 梁国栋,Email:gdliang@hotmail.com
  • 作者简介:刘红,女,博士,讲师,主要从事病毒分子生物学与生物信息学,Email:liuhongseminar@126.com
  • 基金资助:

    国家自然科学基金重大项目(81290342);国家自然科学基金(81301479,81501757);山东省自然科学基金(ZR2013HQ008);传染病预防和控制重点实验室发展项目(2014SKLID103)

The genotype A specific primers for amplifying and sequencing the whole genome of Banna virus

LIU Hong1,2, LIANG Guo-dong2   

  1. 1. School of Life Sciences, Shandong University of Technology, Zibo 255000, Shandong Province, China;
    2. Department of Viral Encephalitis, State Key Laboratory of Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention
  • Received:2016-07-21 Online:2016-12-20 Published:2016-12-20
  • Supported by:

    Supported by the of National Natural Science Foundation of China(Major Program) (No. 81290342), the National Natural Science Foundation of China(No. 81301479, 81501757), the Natural Science Foundation of Shandong Province of China (No. ZR2013HQ008) and the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control (No. 2014SKLID103)

摘要:

目的 设计版纳病毒型别特异性测序引物,为构建快速高效经济的版纳病毒全基因组测序平台奠定基础。方法 根据GenBank已知的版纳病毒基因序列信息进行基因型分析,采用Primer Premier v6.0及Oligo 7.56软件完成版纳病毒亚型特异性扩增引物的设计与评估。使用中国分离的30株版纳病毒进行版纳病毒型别特异性引物工作效率验证和评估。结果 通过对30株隶属于2个不同基因亚型的版纳病毒开展全基因组序列测定,获得2套型别工作效率高、扩增效果稳定的基因A型特异性版纳病毒全基因组序列扩增测序引物。基因A1亚型引物一套共26对;基因A2亚型引物一套共计30对。完成了30株版纳病毒全基因组序列扩增。结论 设计的版纳病毒型别特异性扩增测序引物可以高效准确地完成版纳病毒全基因组序列扩增,为进一步深入研究版纳病毒分子生物学特征奠定了基础。

关键词: 版纳病毒, 基因型, 测序

Abstract:

Objective In order to construct fast and efficient genome sequencing platform of Banna virus (BAV), we designed specific-type sequencing primers for BAV. Methods In this study, primer design software Primer Premier v6.0 was used to design the BAV genotype A specific amplification and sequencing primers to analyze the genotype, the sequences of which were based on the information of BAV on GenBank. Meanwhile, the whole-genome sequences of type A1 and A2 of 30 strains of BAV isolated in China, were sequenced by these primers. Results Two sets of specific amplification and sequencing primers for BAV were designed, 26 pairs for gene A1 subtype, and 30 pairs for gene A2 subtype. Moreover, the whole-genome sequences of 30 strains of BAV were amplified. Conclusion Specific amplification primers of BAV with high efficiency and accuracy were designed, laying foundation for further studies on biological feature of BAV.

Key words: Banna virus, Genotype, Sequencing

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