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박종석(Jong Sug Park),배신철(Shin Chyul Bae),김영민(Young Min Kim),백융기(Young Ki Paik),김주곤(Ju Kon Kim),황영수(Young Soo Hwang) 한국응용생명화학회 1994 Applied Biological Chemistry (Appl Biol Chem) Vol.37 No.3
Rice black-streaked dwarf virus (RBSDV), a member of the plant reoviridae fijivirus group, causes a serious damage for rice production in Korea. To characterize the RBSDV genome, virus particles were produced by feeding of planthopper (Laodelphax striatellus F.) earring RBSDV to maize plants for 2 days. In 30∼40 days after feeding, the viral particles were purified from the infected maize roots by using 10∼40% sucrose gradient centrifugation. After treatment of 10% SDS to remove the viral coat proteins, ten viral double-stranded RNAs were resolved in agrose gel electrophoresis. Total dsRNA was then used to synthesize cDNA by reverse transcriptase and a cDNA library was constructed in the λgt11 vector. The phages that contain RBSDV cDNA fragments were selected by hybridizing with the random-primed probe prepared from RBSDV dsRNAs. After subcloning of several cDNA fragments into the pUC19 plasmid mector, one clone (pRV3) was chosen for sequencing. The pRV3 clone was shown to be Located on the RBSDV genome fragment No.3 by RNA gel-blot analysis. Sequence analysis of the clone revealed that the pRV3 contains two partial open reading frames.