Table 1

Primers used in the study

Figure 1

Schematic of EPA1 and EPA6 gene knockout Note： A： Construction of the EPA1/6 knockout cassettes. The upstream fragment （up） and downstream fragment （down） of the EPA1 gene， along with the HYR resistance marker， were amplified by PCR. Primers EPA1-up-R and HYR-F （red tails） and HYR-R and EPA1-down-F （blue tails） contained reverse-complementary sequences for fusion PCR assembly. The three fragments （mixed at 1∶1∶1.5 ratio） were fused via PCR using primers EPA1-up-F and EPA1-down-R. B： Homologous recombination mechanism. The knockout cassette shares homologous ends （up and down） with the flanking regions of the EPA1/6 gene in C. glabrata strain ATCC 2001. Using the lithium acetate method， the cassette stochastically replaces the genomic EPA1/6 via homologous recombination， achieving targeted gene disruption.

Figure 2

Electrophoresis results of construction of the Candida glabrata EPA1 and EPA6 gene knockout cassette Note： A. EPA1 upstream fragments（up， 1 013 bp） and EPA1 downstream fragments（down， 455 bp）；EPA6 upstream fragments（up， 403 bp） and EPA6 downstream fragments（down， 471 bp）. B. Screening marker fragment（HYR， 1 956 bp）. C. EPA1 knockout cassette（up-HYR-down， 3 424 bp）；EPA6 knockout cassette（up-HYR-down， 2 830 bp）.

Figure 3

PCR verification of the Δepa1, Δepa6 knockout strain and Candida glabrata strain ATCC 2001 Note: (A) PCR amplification of the EPA1/6-in gene；(B) PCR amplification of the EPA1/6-out gene.

Figure 4

Cell surface hydrophobicity of Δepa1, Δepa6 knockout strain and Candida glabrata strain ATCC 2001

Figure 5

Analysis of biofilm formation ability in Candida glabrata strains ATCC 2001, knockout strain Δepa1 and Δepa6