Molecular analysis of the poly-telomeric CTA2 gene family of Candida albicans: Elucidation of genetic or epigenetic regulation of their differential expression, localisation and functional analyses in vitro and during the early phase of colonisation on po

白色念珠菌多端粒 CTA2 基因家族的分子分析：阐明其差异表达的遗传或表观遗传调控、体外定位和功能分析以及定植早期阶段

• 批准号: 5426768
• 负责人: Professor Dr. Jürgen Wendland
• 金额: --
• 依托单位: Institut Mikrobiologie und Biochemie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5426768?language=en
• 关键词: Molecular; analysis; poly; telomeric; CTA2

C. albicans is an opportunistic fungal pathogen that is frequently found in the normal gastrointestinal flora of humans. In immuno-compromised patients or in patients with an altered gut flora, e.g. due to antibiotic treatment, colonization and invasion by C. albicans may occur. Particularly infection of the blood stream by C. albicans (candidaemia) often causes death. We have established an in vitro test system to study the first steps during colonization of C. albicans on the molecular level. We use porcine intestinal epithelium (PIE) of freshly slaughtered pigs, which is inoculated with preincubated C. albicans strains. Under these conditions wildtype cells produce germ tubes after 3h which adhere to the PIE. Cawal1 mutant cells, which under standard laboratory conditions do not form hyphae, show at least adhesion to the epithelium. Using the DNA-array technology (based on the Eurogentec DNA-arrays) genes that are differentially expressed during these early stages of infection (e.g. as compared to a efg1/cph1 null strain) and thus contribute to adhesion and/or hyphal induction will be identified. These differentially expressed genes will be functionally analyzed using PCR-based methods. This includes the generation of deletion strains as well as the subcellular localization of the proteins via GFP-fusions.

C.白色念珠菌是一种常见于人类正常胃肠道植物群中的机会致病真菌。在免疫功能低下的患者或肠道植物群改变的患者中，例如由于抗生素治疗、C.可能出现白色念珠菌。尤其是C.白色念珠菌（念珠菌血症）经常导致死亡。我们建立了一个体外试验系统来研究C.白色念珠菌的分子水平。我们使用新鲜屠宰的猪的猪肠上皮（PIE），其接种有预孵育的C。白色念珠菌菌株。在这些条件下，野生型细胞在3小时后产生芽管，其粘附到PIE。在标准实验室条件下不形成菌丝的Cawal 1突变细胞至少显示出对上皮的粘附。使用DNA阵列技术（基于Eurogentec DNA阵列），将鉴定在这些感染早期阶段差异表达的基因（例如，与efg 1/cph 1无效菌株相比），从而有助于粘附和/或菌丝诱导。将使用基于PCR的方法对这些差异表达的基因进行功能分析。这包括缺失菌株的产生以及通过GFP融合的蛋白质的亚细胞定位。