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

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．