Uhl M Andrew, Johnson Alexander D
Department of Microbiology and Immunology, University of California, San Francisco, CA 94143-0414, USA1.
Microbiology (Reading). 2001 May;147(Pt 5):1189-1195. doi: 10.1099/00221287-147-5-1189.
The study of gene regulation in many organisms has been facilitated by the development of reporter genes. The authors report the use of lacZ from Streptococcus thermophilus, a gene encoding a beta-galactosidase, as a reporter for the fungal pathogen Candida albicans. As test cases, Strep. thermophilus lacZ was placed under control of three different C. albicans promoters: MAL2 (maltase), inducible by maltose; HWP1 (hyphal cell wall protein), induced by conditions that promote filamentous growth; and ACT1 (actin). These constructs were each integrated into the C. albicans genome and beta-galactosidase activity was readily detected from these strains, but only under the appropriate growth conditions. Beta-galactosidase activity could be detected by several methods: quantitative liquid assays using permeabilized cells, colorimetric assays of colonies replicated to paper filters, and in situ coloration of colonies growing on medium containing the indicator X-Gal. These results show the usefulness of STREP: thermophilus lacZ as a monitor of gene regulation in this medically important yeast.
报告基因的发展促进了许多生物体中基因调控的研究。作者报道了将嗜热链球菌的lacZ(一种编码β-半乳糖苷酶的基因)用作真菌病原体白色念珠菌的报告基因。作为测试案例,嗜热链球菌lacZ置于三种不同的白色念珠菌启动子控制之下:MAL2(麦芽糖酶),可被麦芽糖诱导;HWP1(菌丝细胞壁蛋白),在促进丝状生长的条件下被诱导;以及ACT1(肌动蛋白)。这些构建体分别整合到白色念珠菌基因组中,并且很容易从这些菌株中检测到β-半乳糖苷酶活性,但仅在适当的生长条件下。β-半乳糖苷酶活性可以通过几种方法检测:使用透化细胞的定量液体测定、对复制到滤纸的菌落进行比色测定以及对在含有指示剂X-Gal的培养基上生长的菌落进行原位染色。这些结果表明嗜热链球菌lacZ作为这种医学上重要的酵母中基因调控监测指标的有用性。