Yeast vectors for integration at the HO locus.
We have constructed new yeast vectors for targeted integration of desired sequences at the Saccharomyces cerevisiae HO locus. Insertion at HO has been shown to have no effect on yeast growth, and thus these integrations should be neutral. One vector contains the KanMX selectable marker, and integrants can be selected by resistance to G418. The other vector contains the hisG-URA3-hisG cassette, and integrants can be selected by uracil prototrophy. Subsequent growth on 5-FOA permits identification of colonies where recombination between the hisG tandem repeats has led to loss of the URA3 marker and return to uracil auxotrophy. We also describe several new bacterial polylinker vectors derived from pUC21 (ampicillin resistance) and pUK21 (kanamycin resistance).
Voth
W P
WP
Department of Pathology and Department of Biology, University of Utah, Salt Lake City, UT 84132, USA.
Richards
J D
JD
Shaw
J M
JM
Stillman
D J
DJ
D000668:Ampicillin Resistance; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005819:Genetic Markers / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology; D007613:Kanamycin Resistance; D008969:Molecular Sequence Data; D009963:Orotic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D011995:Recombination, Genetic / Q000235:genetics*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins; D020080:Tandem Repeat Sequences / Q000235:genetics
D004269:DNA, Bacterial; D004271:DNA, Fungal; D005656:Fungal Proteins; D005819:Genetic Markers; D005839:Gentamicins; D029701:Saccharomyces cerevisiae Proteins; D009963:Orotic Acid; C001242:5-fluoroorotic acid; C010680:antibiotic G 418; C530845:HO protein, S cerevisiae; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific