DETECTION OF VIROID OF POTATO STREPTOMYCES SCABIES AND PROBE FOR ITS DETECTION

PURPOSE: To detect the potato Streptomyces scabies viroid(PSTVd) with high sensitivity for preventing potatoes from being infected with the viroids by using a plurality of synthetic oligonucleotides which are complementary to the PSTVd RNA as probes. CONSTITUTION: Synthetic oligonucleotides having b...

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Hauptverfasser:	NAKAHARA KENJI, KIMURA IKUO, HATAYA TATSUJI, HAYASHI YOSHINORI, SUGIMOTO NOBUTAKA, YOMO EISHIRO
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA DERIVATIVES THEREOF ENZYMOLOGY INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING NUCLEIC ACIDS NUCLEOSIDES NUCLEOTIDES ORGANIC CHEMISTRY PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS SUGARS TESTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	PURPOSE: To detect the potato Streptomyces scabies viroid(PSTVd) with high sensitivity for preventing potatoes from being infected with the viroids by using a plurality of synthetic oligonucleotides which are complementary to the PSTVd RNA as probes. CONSTITUTION: Synthetic oligonucleotides having bases sequences represented by formulas I to V which are complementary to the RNA of PSTVd are used in a mixture as probes. Raw leaves of tomato or the like infected with PSTVd are ground to extract the nucleic acid soluble in 2M lithium chloride, a reverse transcriptase is allowed to act on to synthesize cDNA, which is amplified by PCR. The amplified cDNA is immobilized to the microplate and a mixture of biotin-labeled synthetic oligonucleotides of formulas I to V is hybridized to the cDNA. Then, avidinated alkali phosphatase is allowed to react with the hybridomas and a color-developing substrate is allowed to act on the reaction mixture to determine PSTVd colorimetrically using a spectrophotometer.