(A) Deletion of
CST20 in C. albicans. PCR with the
divergent oligodeoxynucleotides ODH68 and ODH69 was used to partially
delete the coding sequence of CST20. A
hisG-URA3-hisG cassette was then inserted.
(B) Southern blot analysis with a CST20
fragment from EcoRI–XbaI as a probe. The
genomic DNA samples digested with XhoI were from
following strains: Lanes: 1, CAI4 (ura3/ura3
CST20/CST20); 2, CDH15 (ura3/ura3
CST20/cst20Δ::hisG-URA3-hisG); 3,
CDH18 (ura3/ura3
CST20/cst20Δ::hisG); 4, CDH22
(ura3/ura3
cst20Δ::hisG-URA3hisG/cst20Δ::hisG);
and 5, CDH25 (ura3/ura3
cst20Δ::hisG/cst20Δ::hisG).
(C) Deletion of HST7 in C.
albicans. A HindIII–BglII
fragment was replaced by the hisG-URA3-hisG cassette to
create plasmid pPF-7. (D) Southern blot analysis was
performed with the two large BamHI fragments of pPF-7 as
probes recognizing the HST7 sequence upstream of the
HindIII site and the hisG-URA3-hisG
cassette. The genomic DNA samples digested with XbaI
were from following strains. Lanes: 1, CAI4 (ura3/ura3
HST7/HST7); 2, CDH5 (ura3/ura3
HST7/hst7Δ::hisG-URA3-hisG); 3,
CDH8 (ura3/ura3
HST7/hst7Δ::hisG); 4, CDH10
(ura3/ura3
hst7Δ::hisG-URA3-hisG/hst7Δ::hisG);
and 5, CDH12 (ura3/ura3
hst7Δ::hisG/hst7Δ::hisG).
The endonuclease restriction sites are as follows: B,
BamHI; Bg, BglII; E,
EcoRI; H, HindIII; K,
KpnI; N, NsiI; P, PstI;
Pv, PvuII; Sac, SacI; Sal,
SalI; X, XbaI; Xh, XhoI.