How is it that microsatellites and random oligonucleotides uncover DNA fingerprint patterns?
Minisatellites, microsatellites, and short random oligonucleotides all uncover highly polymorphic DNA fingerprint patterns in Southern analysis of genomic DNA that has been digested with a restriction enzyme having a 4-bp specificity. The polymorphic nature of the fragments is attributed to tandem repeat number variation of embedded minisatellite sequences. This explains why DNA fingerprint fragments are uncovered by minisatellite probes, but does not explain how it is that they are also uncovered by microsatellite and random oligonucleotide probes. To clarify this phenomenon, we sequenced a large bovine genomic BamHI restriction fragment hybridizing to the Jeffreys 33.6 minisatellite probe and consisting of small and large Sau3A-resistant subfragments. The large Sau3A subfragment was found to have a complex architecture, consisting of two different minisatellites, flanked and separated by stretches of unique DNA. The three unique sequences were characterized by sequence simplicity, that is, a higher than chance occurrence of tandem or dispersed repetition of simple sequence motifs. This complex repetitive structure explains the absence of Sau3A restriction sites in the large Sau3A subfragment, yet provides this subfragment with the ability to hybridize to a variety of probe sequences. It is proposed that a large class of interspersed tracts sharing this complex yet simplified sequence structure is found in the genome. Each such tract would have a broad ability to hybridize to a variety of probes, yet would exhibit a dearth of restriction sites. For each restriction enzyme having 4-bp specificity, a subclass of such tracts, completely lacking the corresponding restriction sites, will be present. On digestion with the given restriction enzyme, each such tract would form a large fragment.(ABSTRACT TRUNCATED AT 250 WORDS)
Kashi
Y
Y
Department of Genetics, Alexander Silberman Life Sciences Institute, Hebrew University of Jerusalem, Israel.
Nave
A
A
Darvasi
A
A
Gruenbaum
Y
Y
Soller
M
M
Beckmann
J S
JS
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002417:Cattle; D016172:DNA Fingerprinting; D017931:DNA Primers; D004276:DNA, Satellite / Q000737:chemistry* / Q000235:genetics; D015245:Deoxyribonuclease BamHI; D015252:Deoxyribonucleases, Type II Site-Specific; D014644:Genetic Variation; D015698:Genomic Library; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D011897:Random Allocation; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping
D017931:DNA Primers; D004276:DNA, Satellite; D015345:Oligonucleotide Probes; D015245:Deoxyribonuclease BamHI; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases