» Articles » PMID: 14648114

High-efficiency Transformation of the Pathogenic Yeast Candida Parapsilosis

Overview
Journal Curr Genet
Specialty Genetics
Date 2003 Dec 3
PMID 14648114
Citations 10
Authors
Affiliations
Soon will be listed here.
Abstract

A recently developed transformation system for the pathogenic yeast Candida parapsilosis opened a venue for studying the biological phenomena of this species at the molecular level. However, the standard chemical method yielded only about 1x10(3) transformants/microg of DNA, which is insufficient for certain types of experiment. With the aim of increasing the transformation efficiency, we employed two alternative methods for the introduction of plasmids into the recipient cells. Whereas biolistics resulted in about 5x10(2) transformants/microg of plasmid DNA, electroporation was an order of magnitude more efficient than the chemical method. Pretreatment of cells with 100 mM lithium acetate or 10 mM dithiothreitol resulted in a 5-fold (5x10(4)) or a 10-fold (1x10(5)) increase in transformation efficiency, respectively. This high-efficiency transformation method should be suitable for experiments such as the screening of DNA libraries.

Citing Articles

Chromosome-level genome assembly of an auxotrophic strain of the pathogenic yeast .

Mutalova S, Hodorova V, Brazdovic F, Cillingova A, Tomaska L, Brejova B Microbiol Resour Announc. 2024; 13(9):e0034724.

PMID: 39083682 PMC: 11385725. DOI: 10.1128/mra.00347-24.


Genetic Manipulation as a Tool to Unravel Species Complex Virulence and Drug Resistance: State of the Art.

Zoppo M, Poma N, Di Luca M, Bottai D, Tavanti A J Fungi (Basel). 2021; 7(6).

PMID: 34200514 PMC: 8228522. DOI: 10.3390/jof7060459.


Enhancing the chemical transformation of .

Nemeth T, Nosanchuk J, Vagvolgyi C, Gacser A Virulence. 2021; 12(1):937-950.

PMID: 33729086 PMC: 7993187. DOI: 10.1080/21505594.2021.1893008.


Eukaryotic transporters for hydroxyderivatives of benzoic acid.

Cillingova A, Zeman I, Toth R, Nebohacova M, Dunckova I, Holcova M Sci Rep. 2017; 7(1):8998.

PMID: 28827635 PMC: 5566891. DOI: 10.1038/s41598-017-09408-6.


Generation of Fluorescent Protein Fusions in Candida Species.

Gonia S, Berman J, Gale C J Vis Exp. 2017; (121).

PMID: 28287596 PMC: 5408948. DOI: 10.3791/55333.


References
1.
Voronovsky A, Abbas C, Fayura L, Kshanovska B, Dmytruk K, Sybirna K . Development of a transformation system for the flavinogenic yeast Candida famata. FEMS Yeast Res. 2003; 2(3):381-8. DOI: 10.1016/S1567-1356(02)00112-5. View

2.
Kondo K, Saito T, KAJIWARA S, Takagi M, Misawa N . A transformation system for the yeast Candida utilis: use of a modified endogenous ribosomal protein gene as a drug-resistant marker and ribosomal DNA as an integration target for vector DNA. J Bacteriol. 1995; 177(24):7171-7. PMC: 177597. DOI: 10.1128/jb.177.24.7171-7177.1995. View

3.
Takagi M, Kawai S, Chang M, Shibuya I, Yano K . Construction of a host-vector system in Candida maltosa by using an ARS site isolated from its genome. J Bacteriol. 1986; 167(2):551-5. PMC: 212924. DOI: 10.1128/jb.167.2.551-555.1986. View

4.
Yehuda H, Droby S, Wisniewski M, Goldway M . A transformation system for the biocontrol yeast, Candida oleophila, based on hygromycin B resistance. Curr Genet. 2002; 40(4):282-7. DOI: 10.1007/s00294-001-0255-x. View

5.
Johnston S, Anziano P, Shark K, Sanford J, Butow R . Mitochondrial transformation in yeast by bombardment with microprojectiles. Science. 1988; 240(4858):1538-41. DOI: 10.1126/science.2836954. View