Premium
DNA transformations of Candida tropicalis with replicating and integrative vectors
Author(s) -
Sanglard Dominiqaue,
Fiechter Armin
Publication year - 1992
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320081209
Subject(s) - biology , candida tropicalis , plasmid , selectable marker , transformation (genetics) , autonomously replicating sequence , shuttle vector , genetics , gene , escherichia coli , candida albicans , locus (genetics) , mutant , origin of replication , vector (molecular biology) , recombinant dna
The alkane‐assimilating yeast Candida tropicalis was used as a host for DNA transformations. A stable ade2 mutant (Ha900) obtained by UV‐mutagenesis was used as a recipient for different vectors carrying selectable markers. A first vector, pMK 16, that was developed for the transformation of C. albicans and caries an ADE2 gene marker and a Canadida autonomously replicating sequence (CARS) element promoting autonomous replication, was compatible for transforming Ha900. Two transformant types were observed: (i) pink transformants which easily lose pMK 16 under non‐selective growth conditions; (ii) white transformants, in which the same plasmid exhibited a higher mitotic stability. In both cases pMK 16 could be rescued from these cells in Escherichia coli . A second vector, pADE2, containing the isolated C. tropicalis ADE2 , gene was used to transform Ha900. This vector integrated in the yeast genome at homologous sites of the ade2 locus. Different integration types were observed at one or both ade2 alleles in single or in tandem repeats.