Reporter genes are useful tools to study gene transcription in various organisms. For example, the lacZ gene encoding β-galactosidase has been extensively used as a reporter in bacteria, budding yeast, fruit fly, mouse etc. However, use of this gene in the human fungal pathogen Candida albicans has been limited, probably due to low β-galactosidase activity. Here, we describe a reporter derived from the Vibrio cholerae lacZ gene in which codons have been optimized for expression in C. albicans. The constitutively active ACT1 promoter was fused to this synthetic lacZ reporter and integrated in the C. albicans genome. High β-galactosidase activity in liquid assays was observed for this reporter as well as coloration on X-gal plates. When the lacZ reporter expression was driven by the MET3 promoter, β-galactosidase activity in liquid assays and coloration on X-gal plates was higher in the absence of methionine, thus recapitulating the regulation of the native MET3 gene. This synthetic lacZ gene extends the toolbox of C. albicans reagents by providing a useful reporter for analysis of promoter activity in this organism of medical importance.

中文翻译：

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在人类真菌病原体白色念珠菌中具有高活性的 lacZ 报告基因

报告基因是研究各种生物体中基因转录的有用工具。例如，编码 β-半乳糖苷酶的lacZ基因已被广泛用作细菌、芽殖酵母、果蝇、小鼠等的报告基因。 然而，该基因在人类真菌病原体白色念珠菌中的应用受到限制，可能是由于低β-半乳糖苷酶活性。在这里，我们描述了一种源自霍乱弧菌 lacZ基因的报告基因，其中的密码子已针对在白色念珠菌中的表达进行了优化。组成型活性ACT1启动子与合成的lacZ报告基因融合并整合到白色念珠菌中基因组。在液体测定中观察到该报告基因以及 X-gal 板上的着色具有高 β-半乳糖苷酶活性。当lacZ报告基因表达由MET3启动子驱动时，液体测定中的 β-半乳糖苷酶活性和 X-gal 板上的着色在没有甲硫氨酸的情况下更高，因此概括了天然MET3基因的调节。这种合成的lacZ基因扩展了白色念珠菌试剂的工具箱，为分析这种具有医学重要性的生物体中的启动子活性提供了有用的报告基因。