title: Selection of specific gene probes by combined use of low-stringency PCR amplification and Southern-blot hybridization.
abstract: We have developed a protocol to isolate a gene from which only limited (amino-acid) sequence information is available. It involves two PCR amplifications using one constant primer and a set of nested primers and subsequent crosswise Southern hybridization. The amplified DNA giving a signal in both lanes is further processed for use in gene bank screening by applying standard procedures. In this way the structural gene for a thiol protease, BLH1, the homologue of the bleomycin A (a cancerostatic drug) resistance gene of rabbit (and man), was isolated from yeast genomic DNA.
authors.affiliation: Institut für Genetik und Mikrobiologie der LMU, München, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Magdolen
authors.forename: V
authors.initials: V
authors.lastname: Magdolen
authors.forename: M
authors.initials: M
authors.lastname: Schmitt
authors.forename: W
authors.initials: W
authors.lastname: Bandlow
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods; D015342:DNA Probes / Q000235:genetics*; D015723:Gene Library; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015003:Yeasts / Q000235:genetics*
chemical_list: D015342:DNA Probes