Srikantha T, Klapach A, Lorenz W W, Tsai L K, Laughlin L A, Gorman J A, Soll D R
Department of Biological Sciences, University of Iowa, Iowa City 52242, USA.
J Bacteriol. 1996 Jan;178(1):121-9. doi: 10.1128/jb.178.1.121-129.1996.
The infectious yeast Candida albicans progresses through two developmental programs which involve differential gene expression, the bud-hypha transition and high-frequency phenotypic switching. To understand how differentially expressed genes are regulated in this organism, the promoters of phase-specific genes must be functionally characterized, and a bioluminescent reporter system would facilitate such characterization. However, C. albicans has adopted a nontraditional codon strategy that involves a tRNA with a CAG anticodon to decode the codon CUG as serine rather than leucine. Since the luciferase gene of the sea pansy Renilla reinformis contains no CUGs, we have used it to develop a highly sensitive bioluminescent reporter system for C. albicans. When fused to the galactose-inducible promoter of GAL1, luciferase activity is inducible; when fused to the constitutive EF1 alpha 2 promoter, luciferase activity is constitutive; and when fused to the promoter of the white-phase-specific gene WH11 or the opaque-phase-specific gene OP4, luciferase activity is phase specific. The Renilla luciferase system can, therefore, be used as a bioluminescent reporter to analyze the strength and developmental regulation of C. albicans promoters.
感染性酵母白色念珠菌经历两个发育程序,其中涉及差异基因表达、芽-菌丝转变和高频表型转换。为了解差异表达基因在该生物体中是如何被调控的,必须对阶段特异性基因的启动子进行功能表征,而生物发光报告系统将有助于这种表征。然而,白色念珠菌采用了一种非传统的密码子策略,该策略涉及一种带有CAG反密码子的tRNA,将密码子CUG解码为丝氨酸而非亮氨酸。由于海肾Renilla reinformis的荧光素酶基因不含CUG,我们利用它开发了一种用于白色念珠菌的高灵敏度生物发光报告系统。当与GAL1的半乳糖诱导型启动子融合时,荧光素酶活性是可诱导的;当与组成型EF1α2启动子融合时,荧光素酶活性是组成型的;当与白色阶段特异性基因WH11或不透明阶段特异性基因OP4的启动子融合时,荧光素酶活性是阶段特异性的。因此,海肾荧光素酶系统可以用作生物发光报告物,以分析白色念珠菌启动子的强度和发育调控。