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. 2025 Jan;39(2):e25144.
doi: 10.1002/jcla.25144. Epub 2024 Dec 27.

Construction of Candida albicans pRB895-SAP2-SC5314 With SAP2 High Expression and Its Effects on Adhesion

Lan Xue  1 , Lu Yang  1 , Bingqian Zhao  1 , Wenli Feng  1 , Jing Yang  1 , Yan Ma  1
Affiliations

Construction of Candida albicans pRB895-SAP2-SC5314 With SAP2 High Expression and Its Effects on Adhesion

Lan Xue et al. J Clin Lab Anal. 2025 Jan.

Abstract

Background: SAP2 is closely associated with the pathogenicity and drug resistance of Candida albicans (C. albicans). Our study aimed to construct C. albicans with SAP2 overexpression (pRB895-SAP2-SC5314) to explore the influence of SAP2 on the adhesion of C. albicans and predict the interaction between magnolol and Sap2 by molecular docking.

Methods: The recombinant plasmid pRB895-SAP2 with high SAP2 expression was prepared using a plasmid extraction kit and transformed into C. albicans strain SC5314 using an improved lithium acetate conversion method to construct PRB895-SAP2-SC5314. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the expression of adhesion-related genes in the different strains. Molecular docking and visual analysis of magnolol and Sap2 were performed using the CB-DOCK2 platform.

Results: Compared with the control SC5341 and SC5341 transfected with pRB895, SAP2 expression was significantly higher in the pRB895-SAP2-SC5314 strain (p < 0.05). Based on the sequencing and mapping results, the pRB895-SAP2-SC5314 strain was successfully prepared. SAP2 overexpression significantly downregulated ALS1 expression (p < 0.05), whereas ALS3, TEC1, HOG1, PHR1, and TUP1 expression was downregulated in C. albicans (p < 0.05). The optimal docking result for magnolol and Sap2 protein was -8.1 kcal/mol of vina score, which was considered good docking.

Conclusions: SAP2 overexpression may strengthen the adhesion and pathogenicity of C. albicans, and magnolol may act as an Sap2 inhibitor that affects the adhesion of C. albicans.

Keywords: C. albicans; SAP2; SAP2adhesion; magnolol; pRB895‐SAP2‐SC5314 strain.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Identification of plasmids pRB895 and pRB895‐SAP2, as well as strains pRB895‐SC5314 and pRB895‐SAP2‐SC5314. Gel electrophoresis and mapping results of pRB895‐SAP2 plasmids (A) and empty plasmid pRB895 (B). Sequencing and comparison of pRB895‐SC5314 (C) and pRB895‐SAP2‐SC5314 (D).
FIGURE 2
FIGURE 2
Validation of all primers. Gel electrophoresis results of HOG1, TUP1, SAP2, PHR1, TEC1, ALS3, ALS1, and ACT1 in the strains pRB895‐SAP2‐SC5314 (A), pRB895‐SC5314 (B), and the control strain SC5314 (C). Lanes 4, 5, 6, 7, 8, 9, 10, and 11 represent the primers HOG1, TUP1, SAP2, PHR1, TEC1, ALS3, ALS1 and ACT1, respectively, and lane 12 represents the marker (100–2000 kD).
FIGURE 3
FIGURE 3
Expression of SAP2 in SC5314, pRB895‐SC5314, and pRB895‐SAP2‐SC5314 strains measured using qRT‐PCR. *p < 0.05 versus SC5314; # p < 0.05 versus pRB895‐SC5314.
FIGURE 4
FIGURE 4
Effects of SAP2 on the expression of adhesion‐related genes. The expression levels of ALS1, ALS3, HOG1, PHR1, TEC1, and TUP1 in the different strains were determined using qRT‐PCR. *p < 0.05 versus SC5314; # p < 0.05 versus pRB895‐SC5314.
FIGURE 5
FIGURE 5
Interaction between magnolol and Sap2 proteins by molecular docking. (A) After 20 molecular docking cycles, the binding energies showed good results. (B) The structure visualization of the optimal coupling results (−8.1 kcal/mol of vina score).
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