摘要
目的:构建登革2型病毒中国分离株(DEN2-43)的感染性全长cDNA克隆。方法:根据我室测定的DEN2-43株病毒全基因组序列,利用长链RT-PCR及融合PCR技术扩增此病毒基因组全长cDNA分子,并将其克隆至低拷贝载体pW SK29中构建该病毒株的全长cDNA克隆。将此克隆体外转录后,通过电穿孔导入宿主细胞获得恢复病毒。结果与结论:构建的DEN2-43基因组全长cDNA克隆具有感染性,所获得的恢复病毒具有与原型病毒类似的生物学性质及小鼠乳鼠神经毒力,且可稳定传代。该感染性克隆的构建为深入探讨登革病毒的致病机制及研制新型登革疫苗奠定了基础。
Objective ：To construct infectious full-length cDNA clone of dengue virus type 2 isolated in China （DEN2- 43）. Methods： According to the published nucleotide sequence of DEN243, the approximate 1 lkb full-length cDNAs of dengue virus type 2 were amplified by long RT-PCR and overlop-PCR. Full-length cDNA clone was constructed by cloning the full-length cDNA into a low copy number vector pWSK 29 , from which recovered virus D2012 was acquired by transcription in vitro and electroporation. Results and Conclusion： The full-length cDNA clone of DEN243 constructed was infectious, and recovered virus D212 was indistinguishable from DEN243 in biological properties including suckling mice neurovirulence. The full-length cDNA clone lays the basis for elucidating the mechanism of pathogenesis of dengue virus and developing novel vaccine against dengue fever.
出处
《军事医学科学院院刊》
CSCD
北大核心
2006年第2期116-121,共6页
Bulletin of the Academy of Military Medical Sciences
基金
全军"十五"指令课题(01L054)
