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    • 5. 发明授权
    • Method for generating nonpathogenic infectious pancreatic necrosis virus (IPNV) from synthetic RNA transcripts
    • 从合成RNA转录本产生非致病性感染性胰腺坏死病毒(IPNV)的方法
    • US06274147B1
    • 2001-08-14
    • US09282147
    • 1999-03-31
    • Vikram N. VakhariaKun Yao
    • Vikram N. VakhariaKun Yao
    • A61K3912
    • C12N7/00A61K39/00C07K14/005C12N15/86C12N2720/10022C12N2720/10043C12N2720/10061
    • A system for the generation of live, nonpathogenic infectious pancreatic necrosis virus (IPNV), a segmented double-stranded (ds)RNA virus of the Birnavirdae family, using synthetic transcripts derived from cloned DNA has been developed. Independent full-length cDNA clones were constructed which contained the coding and non-coding regions of RNA segments A and B of IPNV, respectively. Segment A was modified to prevent the expression of NS protein. Synthetic RNAs of both segments were produced by in vitro transcription of linearized plasmids with T7 RNA polymerase. Transfection of CHSE cells with combined plus-sense transcripts of both segments generated infectious virus. The development of a system for producing NS protein deficient IPNV will greatly facilitate studies of viral pathogenesis, and the development of live attenuated vaccines for IPNV.
    • 已经开发了使用衍生自克隆DNA的合成转录物产生活的非致病性感染性胰腺坏死病毒(IPNV),Birnavirdae家族的分段双链(ds)RNA病毒的系统。 构建了独立的全长cDNA克隆,其分别含有IPNV的RNA区段A和B的编码区和非编码区。 修饰A段以防止NS蛋白的表达。 通过T7 RNA聚合酶的线性化质粒的体外转录产生两个片段的合成RNA。 用两个片段的组合加正义转录物转染CHSE细胞产生感染性病毒。 开发NS蛋白缺乏型IPNV的系统将大大促进病毒发病机理的研究,以及IPNV减毒疫苗的开发。