Journal of Diagnostics Concepts & Practice >

2026 , Vol. 25 >Issue 01: 78 - 84

DOI: https://doi.org/10.16150/j.1671-2870.2026.01.011

Original articles

Knockout of EPA1 and EPA6 genes in Candida glabrata and their impact on biofilm formation

  • LIU Jinyan ,
  • SONG Yihui ,
  • WANG Luling ,
  • CHEN Kezhi ,
  • ZHAO Juntao ,
  • LV Geng ,
  • XIANG Mingjie
Expand
  • 1. Radioimmunology and Clinical Laboratory, Ruijin Hospital Luwan Branch, Shanghai Jiao Tong University School of Medicine, Shanghai 200020, China
    2. The Shanghai Institute of Hypertension, 2b. Department of Clinical Laboratory, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    3. Department of Laboratory Medicine, Clinical Laboratory Medicine Research Center, West China Hospital, Sichuan University, Sichuan Clinical Research Center for Laboratory Medicine, Chendu 610041, China

Received date: 2025-09-20

  Revised date: 2025-11-04

  Accepted date: 2025-11-19

  Online published: 2026-02-25

Fold

Abstract

Objective To construct Candida glabrata epithelial adhesin EPA1 and EPA6 gene knockout strains and analyze the effects of gene knockout on the biofilm formation of Candida glabrata. Methods Using fusion PCR and homologous recombination techniques, EPA1 and EPA6 gene knockout constructs were generated, with genomic DNA of Candida glabrata strain ATCC 2001 and plasmid DNA carrying the selectable marker hygromycin resistance gene (HyR) as templates. The knockout constructs were transfected into ATCC 2001 using the lithium acetate transfection method to obtain Δepa1 and Δepa6 knockout strains. The virulence phenotypes of the knockout strains were observed through hydrophobicity tests and biofilm formation experiments. Results Compared with the standard strain ATCC 2001, whose hydrophobicity was 91.5%, the hydrophobicity of Δepa1 and Δepa6 obtained in this study decreased to 61.6% and 75.5%, respectively (P<0.05). After incubation at 30℃ for 4, 16, and 24 hours, the biofilm formation abilities of Δepa1 and Δepa6 were significantly lower than those of the standard strain ATCC 2001 (P<0.05). Conclusions The Candida glabrata EPA1 and EPA6 gene knockout strains are successfully constructed, and it is confirmed that the EPA1 and EPA6 genes can enhance strain virulence biofilm formation.

Key words： Candida glabrata; Epithelial adhesin family; Gene knockout; Biofilm

Cite this article

LIU Jinyan , SONG Yihui , WANG Luling , CHEN Kezhi , ZHAO Juntao , LV Geng , XIANG Mingjie . Knockout of EPA1 and EPA6 genes in Candida glabrata and their impact on biofilm formation[J]. Journal of Diagnostics Concepts & Practice, 2026 , 25(01) : 78 -84 . DOI: 10.16150/j.1671-2870.2026.01.011

References

[1] KATSIPOULAKI M, STAPPERS MHT, MALAVIA-JONES D, et al. Candida albicans and Candida glabrata: global priority pathogens[J]. Microbiol Mol Biol Rev, 2024, 88(2):e0002123.
[2] KRCMERY V, BARNESA J. Non-albicans Candida spp. causing fungaemia: pathogenicity and antifungal resistance[J]. J Hosp Infect, 2002, 50(4):243-260.
[3] FRíAS-DE-LEóN M G, HERNáNDEZ-CASTRO R, CONDE-CUEVAS E, et al. Candida glabrata antifungal resistance and virulence factors, a perfect pathogenic combination[J]. Pharmaceutics, 2021, 13(10):1529.
[4] DE GROOT P W, KRANEVELD E A, YIN Q Y, et al. The cell wall of the human pathogen Candida glabrata: differential incorporation of novel adhesin-like wall proteins[J]. Eukaryot Cell, 2008, 7(11):1951-1964.
[5] 王鲁灵, 刘锦燕, 赵珺涛, 等. 光滑念珠菌毒力因子Epa黏附素的研究进展[J]. 中国真菌学杂志, 2024, 19(3): 320-324.
  WANG L L, LIU J Y, ZHAO J T, et al. Research progress on Epa adhesins, the virulence factors of Candida glabrata[J]. Chin J Mycol, 2024, 19(3):320-324.
[6] ZHAO J T, CHEN K Z, LIU J Y, et al. FLO8 deletion leads to decreased adhesion and virulence with down-regulated expression of EPA1, EPA6, and EPA7 in Candida glabrata[J]. Braz J Microbiol, 2022, 53(2):727-738.
[7] WANG L L, HUANG S J, ZHAO J T, et al. Regulatory role of Mss11 in Candida glabrata virulence: adhesion and biofilm formation[J]. Front Cell Infect Microbiol, 2024, 4:13:1321094.
[8] 王钰婷, 刘锦燕, 史册, 等. 白念珠菌ERG3基因敲除及其对耐药性的影响[J]. 上海交通大学学报(医学版), 2020, 40(2):163-170.
  WANG Y T, LIU J Y, SHI C, et al. Knocking out ERG3 gene of Candida albicans and its effect on drug resistanc[J]. J Shanghai Jiao Tong Univ (Med Sci), 2020, 40(2):163-170.
[9] 赵珺涛, 袁捷, 刘锦燕, 等. 运用融合PCR技术敲除光滑念珠菌FLO8基因以及其对EPA黏附素家族表达的影响[J]. 诊断学理论与实践, 2024, 23(4): 398-404.
  ZHAO J T, YUAN J, LIU J Y, et al. Knocking out FLO8 gene of Candida glabrata and its effect on EPA family[J]. J Diagn Concepts Pract, 2024, 23(4): 398-404.
[10] UENO K, UNO J, NAKAYAMA H, et al. Development of a highly efficient gene targeting system induced by transient repression of YKU80 expression in Candida glabrata[J]. Eukaryot Cell, 2007, 6(7):1239-1247.
[11] MéTIVIER K, ZHOU-LI Y, FAIRHEAD C. In vivo CRISPR-Cas 9 expression in Candida glabrata, Candida bracarensis, and Candida nivariensis: A versatile tool to study chromosomal break repair[J]. Yeast, 2024, 41(9):560-567.
[12] SHEN J, GUO W, K?HLER J R. CaNAT1, a heterologous dominant selectable marker for transformation of Candida albicans and other pathogenic Candida species[J]. Infect Immun, 2005, 73(2):1239-1242.
[13] BASSO LR J R, BARTISS A, MAO Y, et al. Transformation of Candida albicans with a synthetic hygromycin B resistance gene[J]. Yeast, 2010, 27(12):1039-1048.
[14] ESSEN L O, VOGT M S, M?SCH H U. Diversity of GPI-anchored fungal adhesions[J]. Biol Chem, 2020, 401(12):1389-1405.
[15] PAULITSCH A H, WILLINGER B, ZSALATZ B, et al. In-vivo Candida biofilms in scanning electron microscopy[J]. Med Mycol, 2009, 47(7):690-696.
[16] GALOCHA M, PAIS P, CAVALHEIRO M, et al. Divergent approaches to virulence in C. albicans and C. glabrata: Two sides of the same coin[J]. Int J Mol Sci, 2019, 20(9):2345.
Options
Outlines

/

〈 〉