Construction of a library of bovine genomic fragments enriched in CpG islands.
A procedure is described to isolate DNA probes from the bovine genome that are enriched in sites for the so-called rare cutter restriction endonucleases. A collection of SacII (CvCGCGG)-Hin-dIII fragments from bovine sperm was established in the plasmid Bluescript. 180 clones were picked at random and analysed for the presence of inserts with sites for the following rare cutters: EagI, BsshII, NarI, MluI, NruI, NaeI: 70% of the clones contained at least 1 site and 5% contained four different such sites. 22.8% had multiple sites for one or more of the rare cutters tested. Sequence analysis for 16 clones confirmed the cloning of DNA with a G+C content and a proportion of CpG vs GpCs indicative of CpG islands.
Dipartimento di Genetica e Microbiologia, Pavia, Italy.
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F
De Rubertis
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Comincini
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P
Leone
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G
Rognoni
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Ferretti
D000818:Animals; D002417:Cattle / Q000235:genetics*; D003001:Cloning, Molecular; D003596:Cytosine / Q000032:analysis; D015342:DNA Probes / Q000302:isolation & purification*; D015252:Deoxyribonucleases, Type II Site-Specific; D015226:Dinucleoside Phosphates / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006147:Guanine / Q000032:analysis; D008297:Male; D017421:Sequence Analysis; D014170:Transformation, Genetic
D003596:Cytosine; D015342:DNA Probes; D015252:Deoxyribonucleases, Type II Site-Specific; D015226:Dinucleoside Phosphates; D006147:Guanine; C015772:CpG dinucleotide