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A recyclable Candida albicans URA3 cassette for PCR product‐directed gene disruptions
Author(s) -
Bryce Wilson R.,
Davis Dana,
Enloe Brian M.,
Mitchell Aaron P.
Publication year - 2000
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/(sici)1097-0061(20000115)16:1<65::aid-yea508>3.0.co;2-m
Subject(s) - ura3 , biology , genetics , gene , mutant
For some time, gene disruptions in Candida albicans have been made with the hisG‐URA3‐hisG (‘Ura‐blaster’) cassette, which can be re‐used in successive transformations of a single strain after homologous excision of URA3 . However, the hisG repeats are too large for efficient PCR amplification of the entire cassette, so it cannot be used for PCR product‐directed gene disruptions. We describe here a gene disruption cassette, URA3‐dpl200 , with 200 bp flanking repeats that permit efficient PCR amplification. After transformation and integration to produce both arg5::URA3‐dpl200 and rim101::URA3‐dpl200 alleles, we find that arg5::dpl200 and rim101::dpl200 segregants, respectively, can be obtained. We have used the cassette to create rim101::dpl200/rim101::URA3‐dpl200 mutants exclusively through PCR product‐directed disruption. Copyright © 2000 John Wiley & Sons, Ltd.