Construction and identification of SHO1 gene knockout strain of Candida albicans

JIA Keran; QIN Lixia; ZHENG Jie; LI Wei; LI Fang; ZHAO Huihai

doi:10.3969/j.issn.1673-3827.2026.01.009

Abstract：Objective High osmotic pressure signaling protein 1(Sho1)is ubiquitous on the cell membrane of fungi,and its structure is relatively conserved,which plays an important role in the pathogenic mechanism of fungi.However,the effect of Sho1 protein on the pathogenic ability of Candida albicans has not been reported.Therefore,establishing a method to construct SHO1 gene knockout strains of C.albicans will provide a theoretical basis for further exploration of the effect of Sho1 protein on the biological characteristics and pathogenicity of C.albicans.Methods SN152(His-/Leu-/Arg-)deficient strain was selected as the parental strain for gene knockout technology.Fusion PCR and homologous complementation of His and Leu plasmids were used to knock out the target gene rapidly and efficiently.The effect of Sho1 protein on the pathogenic ability of C.albicans was initially explored by crystal violet staining,polystyrene well plate model,and Gram staining oil microscope.Results his and fusion PCR-his gene knockout cassettes were constructed.leu and fusion PCR-leu gene knockout cassettes were transformed into SN152 by electroporation.SHO1 knockout strains were verified by PCR,agarose gel electrophoresis and gene sequencing.Our results suggested that SHO1 knockout decreased the ability of biofilm formation.Conclusion SHO1 gene knockout strain of C.albicans is successfully constructed.SHO1 gene knockout reduces the biofilm formation ability and pathogenicity of C.albicans.