Figure 1

The construction of FLO8 gene knockout elements and FLO8 knockout process A: Amplify the upstream fragment (Up), downstream fragment (Down), and NAT screening fragment of FLO8 gene. There are reverse complementary sequences between primer Up R and NAT F, NAT R and Down F for fusion PCR connection; B: Use primers Up F and Down R to fuse the Up, NAT, and Down fragments for PCR; C: The upper and lower ends of the fused knockout component are homologous to the upstream and downstream of the FLO8 gene of Candida glabrata. Due to the principle of DNA homologous recombination, the NAT sequence of the knockout component has a probability of replacing the FLO8 gene.

Table 1

The sequences of the primers

Figure 2

Identification design for FLO8 knockout

Figure 3

Identification of upstream and downstream fragments, screening marker fragments, and knockout components A: FLO8 upstream and downstream fragments: Up (774 bp) and Down (652 bp); B: screening marker fragment: NAT (1 770 bp); C: FLO8 knockout component: Up-NAT-Down (3 155 bp).

Figure 4

Identification of FLO8 gene knockout in flo8Δ strain A: Primer: T-FLO8 F/R; B: Primer: FLO8-KO F/R.

Figure 5

Expression levels of EPA1, EPA6, and EPA7 in ATCC2001 strain and flo8△ strain For each tested gene, the corresponding gene expression level of the ATCC2001 strain was used as the control (set to 1.0). A: The expression level of EPA1, P<0.001; B: The expression level of EPA6, P<0.001; C: The expression level of EPA7, P<0.001.