| 摘要: |
| 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,realtime PCR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realtime RTPCR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。 |
| 关键词: 白念珠菌 IFD6 生物膜 |
| DOI：10.3724/SP.J.1008.2010.0599 |
| 投稿时间：2009-07-31修订日期：2010-04-26 |
| 基金项目:国家自然科学基金(30500628),上海市教育发展基金会晨光计划(2007CG51). |
|
| Overexpression of IFD6 in Candida albicans promotes biofilm formation |
| LI Ying1,2△, LI De-dong2△, WANG Yan2, LIU Ling-cong1, HUANG Hai2, FU Shou-ting1*, JIANG Yuan-ying2* |
| (1. Department of Pharmacology, School of Life Science and Biological Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China; 2. Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China) |
| Abstract: |
| Objective To investigate the effect of IFD6 on biofilmforming ability of C. albicans. Methods We constructed an IFD6overexpressing plasmid and inserted IFD6 intact open reading frame(ORF) under the control of the MET3 promoter in pCaEXP plasmid, which was then used to transform C. albicans CAI4 by lithium acetate method. PCR was used to investigate the in situ integration of IFD6 gene and realtime PCR was used to select strains highly expressing IFD6 gene. XTT assay and confocal scanning laser microscopy were used to investigate the changes of biofilmforming ability before and after IFD6 overexpression. We also investigated the changes of cell surface hydrophobicity(CSH) using relative CSH assay. Results We successfully constructed IFD6overexpressing plasmid as confirmed by restriction enzyme digestion and sequencing; IFD6overexpressing strain was successfully established as confirmed by PCR. Realtime PCR successfully selected a strain highly expressing IFD6 gene. The results of XTT assay and confocal scanning laser microscopy both showed that IFD6overexpressing strain had an enhanced biofilm forming ability and an increased cell surface hydrophobicity. Conclusion Overexpression of IFD6 can enhance the biofilm forming ability of C. albicans by increasing CSH. |
| Key words: Candida albicans IFD6 biofilms |
