Myers K K, Sypherd P S, Fonzi W A
Genprobe, San Diego, CA 92121, USA.
Curr Genet. 1995 Feb;27(3):243-8. doi: 10.1007/BF00326156.
The C. albicans URA3 gene was tested as a reporter of gene expression. An integrating vector was constructed which contained ADE2 as a selectable marker together with a truncated form of URA3 lacking the first three codons. A DNA fragment containing the promoter and the first 90 codons of the C. albicans CEF3 gene was inserted into the unique XhoI site 5' to URA3 in order to provide an in-frame translational fusion. The functionality of the fusion gene was tested following integration of a single copy of the plasmid into the ADE2 locus. The fusion gene was shown to complement a ura3 deletion mutation and to produce orotidine 5'-monophosphate decarboxylase activity (OMP), which is encoded by URA3. Expression of the fusion gene was appropriately regulated by the growth rate and utilized the same transcriptional start sites as the native CEF3 gene. The results demonstrated that URA3 provides a sensitive and versatile reporter gene for use in C. albicans.
白色念珠菌URA3基因作为基因表达的报告基因进行了测试。构建了一个整合载体，其包含ADE2作为选择标记以及缺少前三个密码子的截短形式的URA3。将包含白色念珠菌CEF3基因启动子和前90个密码子的DNA片段插入到URA3 5'端的独特XhoI位点，以提供框内翻译融合。在将单拷贝质粒整合到ADE2基因座后，测试了融合基因的功能。结果表明，融合基因可弥补ura3缺失突变并产生由URA3编码的乳清苷5'-单磷酸脱羧酶活性（OMP）。融合基因的表达受生长速率的适当调节，并利用与天然CEF3基因相同的转录起始位点。结果表明，URA3为白色念珠菌提供了一个灵敏且通用的报告基因。
