RT Journal
A1 Castaño, Irene
A1 Kaur, Rupinder
A1 Pan, Shihjung
A1 Cregg, Robert
A1 Peñas, Alejandro De Las
A1 Guo, Nini
A1 Biery, Matthew C.
A1 Craig, Nancy L.
A1 Cormack, Brendan P.
T1 Tn7-Based Genome-Wide Random Insertional Mutagenesis of Candida glabrata

JF Genome Research 
JO Genome Research 
YR 2003 
FD May 01 
VO 13 
IS 5 
SP 905 
OP 915 
DO 10.1101/gr.848203 
UL http://genome.cshlp.org/content/13/5/905.abstract 
AB We describe and characterize a method for insertional mutagenesis of the yeast pathogen Candida glabrata using the bacterial transposon Tn7. Tn7 was used to mutagenize a C. glabrata genomic fosmid library. Pools of random Tn7insertions in individual fosmids were recovered by transformation intoEscherichia coli. Subsequently, these were introduced by recombination into the C. glabrata genome. We found thatC. glabrata genomic fragments carrying a Tn7insertion could integrate into the genome by nonhomologous recombination, by single crossover (generating a duplication of the insertionally mutagenized locus), and by double crossover, yielding an allele replacement. We were able to generate a highly representative set of ∼104 allele replacements in C. glabrata, and an initial characterization of these shows that a wide diversity of genes were targeted in the mutagenesis. Because the identity of disrupted genes for any mutant of interest can be rapidly identified, this method should be of general utility in functional genomic characterization of this important yeast pathogen. In addition, the method might be broadly applicable to mutational analysis of other organisms.