Development of a set of plasmid vectors for genetic manipulations of the pathogenic yeast Candida parapsilosis.
A system for genetic transformation of the yeast Candida parapsilosis, recently developed in our laboratory, opened a venue for investigation of this pathogenic species at the molecular level. In this study we extend the range of available experimental tools by construction of a genomic DNA library suitable for screening and isolation of genes by functional complementation of yeast mutants and a set of replicative plasmid vectors for genetic manipulation of C. parapsilosis cells. The plasmids are based on auxotrophic (CpGAL1, CpURA3, CpMET2, CpLYS4) and dominant (CaIMH3) selection markers. In addition, we constructed plasmid derivatives containing reporter genes yEGFP3 and KlLAC4 coding for enhanced version of the green fluorescent protein and Kluyveromyces lactis beta-galactosidase, respectively. The vectors facilitate propagation and expression of cloned genes in C. parapsilosis cells and allow intracellular localization of gene products and/or monitoring the activity of promoter sequences.
Kosa
Peter
P
Department of Biochemistry and Genetics, Faculty of Natural Sciences, Comenius University, Mlynska dolina CH-1, 842 15 Bratislava, Slovak Republic. pkosa@fns.uniba.sk
Gavenciakova
Barbora
B
Nosek
Jozef
J
D002175:Candida / Q000235:genetics*; D003001:Cloning, Molecular; D017930:Genes, Reporter; D005816:Genetic Complementation Test; D005821:Genetic Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007716:Kluyveromyces / Q000378:metabolism; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D049452:Green Fluorescent Proteins