The ble gene of Streptoalloteichus hindustanus as a new selectable marker for Dictyostelium discoideum confers resistance to phleomycin.
An expression cassette has been constructed which allows expression of the ble gene isolated from Streptoalloteichus hindustanus in Dictyostelium discoideum. This construct has been shown to confer resistance to the bleomycin related antibiotic phleomycin. since the uptake of phleomycin by the cells is pH dependent, we established conditions that allow selection of phleomycin-resistant transformants. Vectors pfeI and pfeII contain, in addition to the cassette, a 592 bp fragment of the D. discoideum plasmid Ddp2 that enables the plasmids to replicate extrachromosomally in Dictyostelium when transformed into a strain that expresses a Ddp2-specific transacting factor (12). pfeI and pfeII contain various unique restriction enzyme sites for cloning. They differ in the G/C-content of the sequence upstream of the ATG start codon.
Leiting
B
B
Max-Planck-Institut für Biochemie, Martinsried bei München, F.R.G.
Noegel
A A
AA
D000818:Animals; D001420:Bacteria, Aerobic / Q000187:drug effects / Q000235:genetics*; D001483:Base Sequence; D004023:Dictyostelium / Q000235:genetics*; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005798:Genes, Bacterial; D005800:Genes, Fungal; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010692:Phleomycins / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D005656:Fungal Proteins; D009838:Oligodeoxyribonucleotides; D010692:Phleomycins