In vitro destabilization of plant viruses and cDNA synthesis

• Published in: Journal of virological methods Vol. 44; no. 2; pp. 211 - 220
• Main Authors: Wyatt, Stephen D., Druffel, Keri, Berger, Philip H.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.10.1993; Amsterdam Elsevier; New York, NY
• Subjects: ADN; Agronomy. Soil science and plant productions; ARN; bean common mosaic virus; Biological and medical sciences; Blotting, Southern; Bromovirus - chemistry; Bromovirus - genetics; cDNA synthesis; Centrifugation, Density Gradient; cowpea chlorotic mottle virus; Destabilization; DNA; DNA, Complementary - biosynthesis; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; Genome, Viral; Microbiology; Plant virus; PLANT VIRUSES; Plant Viruses - chemistry; Plant Viruses - genetics; Potyvirus - chemistry; Potyvirus - genetics; RNA; RNA, Viral - genetics; Techniques used in virology; Transcription, Genetic; Virology; VIRUS DE LAS PLANTAS; VIRUS DES VEGETAUX; Plant virus; Destabilization; cDNA synthesis; Bromovirus; Virus; Purification; Complementary DNA; Plant pathogen; RNA; Cowpea chlorotic mottle virus; DNA synthesis; Bean common mosaic virus; Potyvirus
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/0166-0934(93)90056-W

DNA copies of a wide range of RNA viruses can be made by the direct addition of appropriately treated, purified virus particles to a reverse transcription reaction. Therefore, many problems associated with RNA isolation can be circumvented. Virus particles can be sufficiently destabilized by adjustments of salt content, buffer, pH or by the use of physical force supplied by a freeze/thaw cycle so that RNA in sufficient quantity and physical condition is available for the synthesis of in some cases, full length cDNAs. cDNAs have been made of viruses in the bromo-, poty-, carla-, ilar-, potex-, tobra and tobamovirus groups. Reported here are experiments with cowpea chlorotic mottle virus and bean common mosaic virus.