Sánchez M, Prado M, Iglesias F J, Domínguez A
Departamento de Microbiología y Genética, Universidad de Salamanca, Spain.
Yeast. 1995 Apr 30;11(5):425-33. doi: 10.1002/yea.320110505.
The URA5 gene of Yarrowia lipolytica encoding the orotate phosphoribosyl transferase (OPRTase, EC2.4.2.10) was isolated by target integration in a mutant strain originally named ura2.21. The nucleotide sequence of the gene predicts a protein with high similarities with the OPRTases from Saccharomyces cerevisiae, Podospora anserina and Escherichia coli and to a lesser extent with that of Dictyostelium discoideum. The transcription start point has been mapped by primer extension analysis and indicates the existence of a long leader sequence in the corresponding mRNA. Northern-blot hybridization revealed the URA5 transcript to be approximately 0.94 kb. Deletion of the URA5 gene in Y. lipolytica produced a leaky phenotype similar to the one described for the ura5 mutation in S. cerevisiae. The URA5 gene of Y. lipolytica was able to complement functionally the ura5 mutation of S. cerevisiae.
通过在最初命名为ura2.21的突变菌株中进行靶向整合，分离出解脂耶氏酵母中编码乳清酸磷酸核糖基转移酶（OPRTase，EC2.4.2.10）的URA5基因。该基因的核苷酸序列预测其编码的蛋白质与酿酒酵母、嗜热栖热放线菌和大肠杆菌的OPRTase具有高度相似性，与盘基网柄菌的OPRTase相似性较低。通过引物延伸分析确定了转录起始点，表明相应mRNA中存在长的前导序列。Northern杂交显示URA5转录本约为0.94 kb。解脂耶氏酵母中URA5基因的缺失产生了一种渗漏表型，类似于酿酒酵母中ura5突变所描述的表型。解脂耶氏酵母的URA5基因能够在功能上互补酿酒酵母的ura5突变。
