Use of URA3 as a reporter of gene expression in C. albicans.
Use of URA3 as a reporter of gene expression in C. albicans.
复制标题
使用 URA3 作为白色念珠菌基因表达的报告基因。
DOI：
10.1007/bf00326156
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发表时间：
1995
期刊：
影响因子：
2.5
通讯作者：
Fonzi,WA
中科院分区：
文献类型：
--
作者：
Myers,KK;Sypherd,PS;Fonzi,WA
C.白念珠菌URA 3基因作为基因表达报告基因进行了检测。构建了一个整合载体，该载体含有作为选择标记的ADE 2以及缺失前三个密码子的URA 3的截短形式。一个含有启动子和C.将白念珠菌CEF 3基因插入URA 3的5′端独特的XhoI位点，以提供符合读框的翻译融合。将单个拷贝的质粒整合到ADE 2基因座中后，检测融合基因的功能性。该融合基因可互补aura 3缺失突变，并产生乳清酸核苷5′-单磷酸脱羧酶（OMP）活性，该活性由URA 3编码。融合基因的表达受生长速率的适当调控，并利用与天然CEF 3基因相同的转录起始位点。结果表明，URA 3为C.白色念珠菌
TheC. albicans URA3gene was tested as a reporter of gene expression. An integrating vector was constructed which containedADE2as a selectable marker together with a truncated form ofURA3lacking the first three codons. A DNA fragment containing the promoter and the first 90 codons of theC. albicans CEF3gene was inserted into the uniqueXhoI site 5′ toURA3in 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 theADE2locus. The fusion gene was shown to complement aura3deletion mutation and to produce orotidine 5′-monophosphate decarboxylase activity (OMP), which is encoded byURA3. Expression of the fusion gene was appropriately regulated by the growth rate and utilized the same transcriptional start sites as the nativeCEF3gene. The results demonstrated thatURA3provides a sensitive and versatile reporter gene for use inC. albicans.