Recombinant DNA probes and polymerase chain reaction for detection of Mycoplasma gallisepticum strains.
Two recombinant DNA clones, pMG286.2 and pMG301.1, were isolated from the partial genomic library of Mycoplasma gallisepticum strain S6. Recombinant M. gallisepticum specific fragments were used as probes in Southern hybridisation with 10 M. gallisepticum strains whose DNA was digested by EcoRI, HindIII, BglII, RsaI and BamHI. The 1.5 kb fragment pMG301.1 did not show polymorphism in hybridisation patterns with M. gallisepticum strains, while the 3.5 kb fragment pMG286.2 enabled differentiation of M. gallisepticum strains into clusters. The DNA sequence of pMG301.1 was used to design a pair of 27-mer oligonucleotides flanking a 1.3 kb genomic region. These two primers directed specific in vitro amplification of all M. gallisepticum strains assayed giving an expected 1.3 kb product. Digestion of polymerase chain reaction products by DdeI enabled simple differentiation between clusters of M. gallisepticum strains and may be useful for improved epizootiological studies of M. gallisepticum infections in poultry.
Dovc
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University of Ljubljana, Biotechnical Faculty, Zootechnical Department, Domzale, Slovenia.
Bencina
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Antes
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Mann
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W
D001483:Base Sequence; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D004274:DNA, Recombinant; D008969:Molecular Sequence Data; D009174:Mycoplasma / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D013045:Species Specificity
D015342:DNA Probes; D004274:DNA, Recombinant