Identification of procaryotic repetitive DNA suitable for use as fingerprinting probes.
We have developed a method which enables the cloning and identification of procaryotic repetitive DNA suitable for use as DNA fingerprinting probes. The method involves shotgun cloning of restricted genomic DNA with subsequent selection of clones containing repetitive DNA by reverse-probed genomic hybridizations, in which the plasmid DNA clones are probed with labelled genomic DNA. Confirmation that the clones contained repeated sequences was by Southern hybridization, gene copy equivalence, and DNA sequencing. The sequences were used for highly specific and sensitive detection of bacteria and as target sequences for the mediation of chromosomal integration of reporter gene constructs.
Waterhouse
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RN
Department of Molecular and Cell Biology, Marischal College, University of Aberdeen, Scotland.
Glover
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LA
D001412:Bacillus subtilis / Q000235:genetics; D015139:Blotting, Southern; D003001:Cloning, Molecular; D016172:DNA Fingerprinting / Q000379:methods*; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D011549:Pseudomonas / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping
D015342:DNA Probes; D004269:DNA, Bacterial