ISSN:
1365-3059
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
Notes:
The naturally polyadenylated genomic RNA of PYV served as a template for the synthesis of complementary DNA (cDNA) strand-primed with oligo-dT using the enzyme avian myeloblastosis virus reverse transcriptase. A second cDNA strand was made with DNA polymerase I and the double-stranded product (about 10 kbp) was cleaved with a Hind III restriction endonuclease. The resulting five main fragments of 800,950, 1250,2400 and 2600 bp were subsequently cloned into the Hind III site of the pBR322 plasmid of Escherichia coli. Three types of cDNA inserts (800, 950 and 2400 bp) were detected among the isolated recombinant plasmid clones. Intact untreated virus particles and phenolized virus samples hybridized with the cloned cDNA sequences. These clones hybridized with a major virus RNA band of about 10 kbp and a minor band of higher molecular weight in blots of RNA extracted from purified virus particles and fractionated in a denaturing agarose gel. The prospects of using cloned PVY sequences for the analysis of the virus genome and for virus detection are discussed.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1365-3059.1986.tb02002.x
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