Rapid Purification and in vitro Translation of Heliothis armigera Nuclear Polyhedrosis Virus mRNA
Abstract: Total RNAs were isolated from Heliothis armigera larvae infected with Heliothis armigeraNuclear Polyhedrosis Virus(HaNPV) late after infection(60 h postinfection),800 μg total RNAswere obtained from 1g larvae by improved granidinium isothiocgarate method.The whole viralDNA was purified from HaNPV polyhedra.The DNA was immobilized on nitrocellulose filtersand used to select viral specific mRNAs from total RNA.The specific mRNAs were translated invitro in the presence of L-(35S)methionine by using a rabbit reticulocyte lysate system and ana-lyzed on sodium dodecyl sulfate-polyacrylamide gel.At least twenty proteins were obtained.Theviral structural proteins were among the translation products which were testified with the experi-ment of ELISA.