Construction and identification of reverse genetics system of Dengue type 2 virus isolated in China

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• 作者：Wuyang Zhu (1)
  Shuiping Chen (1)
  Chenggeng Qin (1)
  Man Yu (1)
  Tao Jiang (1)
  Yongqiang Deng (1)
  Ede Qin (1)
  
• 关键词：dengue virus ; reverse genetics system ; rescued virus
• 刊名：Chinese Science Bulletin
• 出版年：2006
• 出版时间：September 2006
• 年：2006
• 卷：51
• 期：17
• 页码：2065-2071
• 全文大小：7543KB
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• 作者单位：Wuyang Zhu (1)
  Shuiping Chen (1)
  Chenggeng Qin (1)
  Man Yu (1)
  Tao Jiang (1)
  Yongqiang Deng (1)
  Ede Qin (1)
  
  1. State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing, 100071, China
  
• ISSN：1861-9541

文摘

To construct infectious full-length cDNA clone of dengue virus type 2 isolated in China (DEN2-43), according to the published nucleotide sequence of the virus strain, the approximately 11 kb full-length cDNAs of DEN2-43 were amplified by long RT-PCR and fusion PCR. Full-length cDNA clones were constructed by inserting the full-length cDNA into a low copy vector pWSK29, from which rescued virus D212 was acquired by transcription in vitro and electroporation. The full-length cDNA clone pD212 was infectious, and rescued virus acquired in C6/36 cells was indistinguishable from DEN2-43 virus in biological properties including suckling mice neurovirulence. The reverse genetics system helps elucidate the mechanism of pathogenesis of dengue virus and develop novel vaccine against dengue.