Citation: ZHANG Shu—yonga.Bonami Jean-Robert, SHI Zheng-li ”, . cDNA Library Construction of a Chinese mitten crab reovirus RNA1 and Partial Sequence Analysis of its RNA Polymerase Gene .VIROLOGICA SINICA, 2003, 18(1) : 72-75.

cDNA Library Construction of a Chinese mitten crab reovirus RNA1 and Partial Sequence Analysis of its RNA Polymerase Gene

  • Available online: 02 February 2003
  • A reovirus (designated as EsRV905), was isolated from Chinese Mitten Crab(Eriocheir sinensis). Viral genome was extxacted with Trizol reagent and individual segment was recovered separately from gel after 6.5% PAGE. The cDNA library of RNA1 was synthesized using random primer method and short fragments l ess than 200 bp were discarded using gel recovery kit. Th e cDNA was ligated to blunt-end plasmid and transformed to competent cel1. Positive clones were screened by blue and white colonies and checked by enzyme digestion. Two plasmids containing around 1.5kb inserts from the first segment were selected for sequencing. One of them contained the main motif of RNA polymerase(RdRp).The RdRp of EsRV905 is located in the first segment of EsRV905 genome.

  • 加载中
  • 加载中

Article Metrics

Article views(3866) PDF downloads(797) Cited by()

Related
Proportional views

    cDNA Library Construction of a Chinese mitten crab reovirus RNA1 and Partial Sequence Analysis of its RNA Polymerase Gene

    • 1. 1.Joint-lab ofInvertebrate Virology,Wuhan Institute ofVirology,Chinese Academy ofScienees,Wuhan 430071,China
    • 2. UMR5098.DRIM,CNRS/IFREMER/UMII,cc一80,P/ace Eugene Bataillon,34095 Montpellier Cedex5,France

    Abstract: A reovirus (designated as EsRV905), was isolated from Chinese Mitten Crab(Eriocheir sinensis). Viral genome was extxacted with Trizol reagent and individual segment was recovered separately from gel after 6.5% PAGE. The cDNA library of RNA1 was synthesized using random primer method and short fragments l ess than 200 bp were discarded using gel recovery kit. Th e cDNA was ligated to blunt-end plasmid and transformed to competent cel1. Positive clones were screened by blue and white colonies and checked by enzyme digestion. Two plasmids containing around 1.5kb inserts from the first segment were selected for sequencing. One of them contained the main motif of RNA polymerase(RdRp).The RdRp of EsRV905 is located in the first segment of EsRV905 genome.

    Relative (20)

    目录

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return