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Virologica Sinica, 18 (1) : 72-75, 2003
cDNA Library Construction of a Chinese mitten crab reovirus RNA1 and Partial Sequence Analysis of its RNA Polymerase Gene
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
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.
  Published online: 2 Feb 2003
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