title: A novel kanamycin/G418 resistance marker for direct selection of transformants in Escherichia coli and different yeast species.
abstract: We have developed a set of cloning vectors possessing a modified Tn903 kanamycin resistance gene that enables the selection of both kanamycin-resistant transformants in Escherichia coli and G418-resistant transformants in the yeasts Saccharomyces cerevisiae, Hansenula polymorpha and Pichia pastoris. Expression of this gene in yeast is controlled by the H. polymorpha glyceraldehyde-3-phosphate dehydrogenase promoter, while expression in E. coli is governed by an upstream E. coli lacZ promoter. Applicability of the vectors for gene disruption in H. polymorpha and S. cerevisiae was demonstrated by inactivation of the HpMAL1 and URA3 genes, respectively. One of the vectors possesses a H. polymorpha ARS allowing plasmid maintenance in an episomal state. The small size of the vectors (2-2.5 kb) makes them convenient for routine DNA cloning. In addition, we report a novel approach for construction of gene disruption cassettes.
authors.affiliation: Institute of Experimental Cardiology, Cardiology Research Centre, 121552 Moscow, Russia. moagaphonov@cardio.ru
authors.forename: Michael
authors.initials: M
authors.lastname: Agaphonov
authors.forename: Nina
authors.initials: N
authors.lastname: Romanova
authors.forename: Eui-Sung
authors.initials: ES
authors.lastname: Choi
authors.forename: Michael
authors.initials: M
authors.lastname: Ter-Avanesyan
mesh_terms: D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005819:Genetic Markers; D005839:Gentamicins / Q000494:pharmacology*; D007612:Kanamycin / Q000494:pharmacology*; D014170:Transformation, Genetic; D015003:Yeasts / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D005819:Genetic Markers; D005839:Gentamicins; D007612:Kanamycin; C010680:antibiotic G 418