论文部分内容阅读
目的考察白念珠菌IFD6基因对生物被膜形成能力的影响。方法构建白念珠菌IFD6基因高表达质粒,将IFD6基因开放阅读框(ORF)置于pCaEXP高表达载体中MET3启动子的控制下,醋酸锂法转染白念珠菌CAI4,PCR鉴定IFD6基因的原位整合,real-timePCR筛选IFD6基因表达量明显升高的菌株。利用XTT法和激光共聚焦显微镜测定并观察IFD6基因高表达前后生物被膜形成能力的变化,利用细胞表面疏水性实验考察白念珠菌细胞表面疏水性的变化。结果通过酶切和测序验证IFD6基因高表达质粒构建正确;通过PCR验证IFD6基因高表达菌株构建正确,并通过real-timeRT-PCR筛选得到1株IFD6基因表达量相对较高的菌株。XTT法和激光共聚焦显微镜观察的结果一致显示IFD6基因高表达菌形成生物被膜的能力增强,细胞表面疏水性增加。结论 IFD6基因高表达能增加白念珠菌细胞表面疏水性,从而使白念珠菌生物被膜的形成能力增强。
Objective To investigate the effect of Candida albicans IFD6 gene on the biofilm formation ability. Methods The high expression plasmid of IFD6 gene of Candida albicans was constructed. The IFD6 open reading frame (ORF) was placed under the control of MET3 promoter in pCaEXP high expression vector. The Candida albicans CAI4 was transfected by lithium acetate method. Bit integration, real-time PCR screening IFD6 gene expression was significantly higher strains. The changes of biofilm formation ability before and after IFD6 gene expression were measured by XTT method and laser confocal microscopy. The cell surface hydrophobicity was used to investigate the changes of cell surface hydrophobicity of Candida albicans cells. Results The plasmid IFD6 was confirmed by restriction enzyme digestion and sequencing. The IFD6 gene highly expressed was confirmed by PCR and the strain with high IFD6 expression was screened by real-time RT-PCR. XTT method and laser confocal microscopy showed that the high expression of IFD6 gene enhanced the ability of biofilm formation and the cell surface hydrophobicity increased. Conclusion The high expression of IFD6 gene can increase the surface hydrophobicity of Candida albicans cells and enhance the formation of C. albicans biofilm.