Gene mapping of cytoplasmic polyhedrosis virus of silkworm by the full-length mRNA prepared under optimized conditions of transcription in vitro.
Viral mRNA synthesis by the RNA polymerase associated with purified cytoplasmic polyhedrosis virus (CPV) was studied. The formation of full-length mRNA products was facilitated by including in the reaction mixture 100 mM sodium acetate, high concentrations of ribonucleoside triphosphates, and proteinase K. The 10 different species of CPV mRNAS were resolved into 9 discrete RNA bands by agarose gel electrophoresis at pH 3.5 in buffer containing 7 M urea. Each purified viral mRNA hybridized specifically to one of the viral genome segments which were separated by polyacrylamide gel electrophoresis into the 10 species of dsRNA. The relationship between the genome segments and their cognate mRNAs synthesized in vitro is thus established. Under optimal conditions of mRNA synthesis each of the genome segments was transcribed at a similar rate as determined from the yield of individual separated mRNA species. A recycling model of genome-associated RNA polymerase for viral transcription is discussed.
Smith
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RE
Roche Institute of Molecular Biology, Nutley, New Jersey 07110, USA.
Furuichi
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Y