Direct selection of DNA sequences conserved between species.
An essential requirement in the analysis of genomes is the identification of functionally important sequence elements, which are often evolutionarily conserved. We describe here the development of a novel procedure for the selective isolation of conserved sequences which is based on hybridization of PCR-amplifiable DNA fragments from the whole genome of one species to biotinylated DNA from a genomic region of another species. The interspecies DNA hybrids are immobilized and the PCR-amplifiable DNA fragments are eluted, amplified and after further hybridization-amplification rounds cloned. This method was used for the generation of sublibraries of conserved sequences from mouse and pig DNA from regions corresponding to cosmids from the human Xq28 region. Mouse and pig homologs of sequences containing exons of known human genes, as well as exons from novel genes have been identified.
Sedlacek
Z
Z
Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Konecki
D S
DS
Siebenhaar
R
R
Kioschis
P
P
Poustka
A
A
D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D002352:Carrier Proteins / Q000235:genetics; D017124:Conserved Sequence; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D015342:DNA Probes; D015723:Gene Library; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D000080882:Ribosomal Protein L10; D012269:Ribosomal Proteins; D017385:Sequence Homology; D013045:Species Specificity; D013552:Swine
D002352:Carrier Proteins; D015342:DNA Probes; C000630686:RPL10 protein, human; D012269:Ribosomal Proteins; C000630690:Rpl10 protein, mouse; D004247:DNA; D005954:Glucosephosphate Dehydrogenase