[Cloning of Chrysanthium stund virion cDNA in pUC19 plasmid and the use of cloned cDNA for detecting the virion].
26 base long deoxyribonucleotide complementary to the lower part of the Central Conserved Region of chrysanthemum stund viroid (CSV) was used for synthesis of the first strand cDNA. The cDNA was cloned into plasmid vector pUC19 and the primary structure was determined. Cloned, full length cDNA was used as hybridisation probe for detection of CSV. It was possible to detect about 26 pg of purified CSV RNA immobilized on nitrocellulose filters using 32P-labeled probe. In the case of biotinylated probe it was possible to detect about 26 pg of purified CSV RNA visualizing results by streptavidin-alkaline phosphatase conjugates. It has been shown that such a cloned cDNA can be used for wide scale detection of CSV.
Meldraĭs
Ia A
IaA
Line
I E
IE
Ivanova
M N
MN
Vinkele
R A
RA
Druka
A Ia
AIa
Gurinovich
T I
TI
D001483:Base Sequence; D011723:Chrysanthemum cinerariifolium / Q000382:microbiology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010942:Plant Viruses / Q000235:genetics*; D010957:Plasmids; D014771:Virion / Q000235:genetics* / Q000302:isolation & purification
D004279:DNA, Viral; D004247:DNA
Klonirovanie kDNK viroida karlikovosti khrizantem v plazmide pUC19 i ispol'zovanie klonirovannoĭ kDNK dliia obnaruzheniia viroida karlikovosti khrizantem.