Analysis of Mycoplasma hyorhinis DNA in the presence of host cells without growing the mycoplasma axenically.
Mycoplasma hyorhinis coisolates with the mitochondria of the cells in which it is carried as an infection. Since both mitochondria and mycoplasmas synthesize DNA by using the prokaryotic DNA polymerase gamma, the use of aphidicolin, which inhibits eukaryotic DNA polymerase alpha, allows for selective synthesis of mycoplasmal and mitochondrial DNA. The restriction patterns of mitochondria and mycoplasmas can easily be differentiated from each other in mixtures of both DNAs. Thus, it is possible to study the molecular biology of this noncultivable mycoplasma in situ rather than after growth in artificial media, with its potential genetic consequences during adjustment to axenic growth.
Schaeffer
W I
WI
Department of Microbiology and Molecular Genetics, Colleges of Agriculture and Life Sciences, University of Vermont, Burlington 05405.
Olson
L D
LD
Barile
M F
MF
Sun
F W
FW
D000818:Animals; D016590:Aphidicolin; D015139:Blotting, Southern; D002478:Cells, Cultured; D004257:DNA Polymerase II / Q000037:antagonists & inhibitors; D004269:DNA, Bacterial / Q000032:analysis* / Q000302:isolation & purification; D004272:DNA, Mitochondrial / Q000032:analysis; D004224:Diterpenes / Q000494:pharmacology; D066298:In Vitro Techniques; D009174:Mycoplasma / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D051381:Rats
D004269:DNA, Bacterial; D004272:DNA, Mitochondrial; D004224:Diterpenes; D016590:Aphidicolin; D004257:DNA Polymerase II