Cloned riboprobe for detection of a mycoplasmalike organism.
A [32P]-labeled single stranded-RNA probe (riboprobe) was constructed with plasmid vector pSP64 and used to detect and specifically identify an uncultured pathogenic mycoplasmalike organism in infected host. The riboprobe was more sensitive and reliable than complementary double stranded-DNA probe in detection of western X mycoplasmalike organism. When concentration of a double stranded-DNA probe was increased, nonspecific hybridization signal was observed with nucleic acid from healthy plants and from plants infected by other mycoplasmalike organisms. In contrast, sensitivity of detection with the complementary riboprobe was increased at elevated probe concentrations without nonspecific hybridization.
Lee
I M
IM
Microbiology and Plant Pathology Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705.
Davis
R E
RE
Hammond
R
R
Kirkpatrick
B
B
D003001:Cloning, Molecular / Q000379:methods; D003101:Collodion; D004262:DNA Restriction Enzymes; D004591:Electrophoresis, Polyacrylamide Gel; D009174:Mycoplasma / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D010935:Plant Diseases; D012313:RNA
D012313:RNA; D003101:Collodion; D004262:DNA Restriction Enzymes