A neutral trehalase gene from Candida albicans:: molecular cloning, characterization and disruption
A neutral trehalase gene from Candida albicans:: molecular cloning, characterization and disruption
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DOI：
10.1099/00221287-143-12-3747
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发表时间：
1997-12-01
期刊：
影响因子：
--
通讯作者：
Künkel, W
中科院分区：
文献类型：
--
作者：
Eck, R;Bergmann, C;Künkel, W
中性海藻糖酶基因。从人类致病性酵母菌白色念珠菌中分离并鉴定了NTC 1。该基因的开放阅读框为2724 bp，编码907个氨基酸，分子量为104 kDa。通过北方印迹分析检测到约3.2 kb的单个转录物。将C.白色念珠菌NTC 1基因产物与酿酒酵母NTH 1基因产物的同源性为57%。NTC 1基因定位于1号染色体或R.通过序列基因破坏构建了一个无效突变体（Delta ntc 1/Delta ntc 1）。中性海藻糖酶缺失纯合突变体的提取物只有边际中性海藻糖酶活性。杂合突变体的提取物显示出野生型菌株和纯合突变体的提取物之间的中间活性。在小鼠系统性念珠菌病模型中，与野生型菌株相比，无效突变株的致病性没有显着差异。这一结果表明，中性海藻糖酶C。白色念珠菌不是抗真菌药物的潜在靶点。
A neutral trehalase gene. NTC1, from the human pathogenic yeast Candida albicans was isolated and characterized. An ORF of 2724 bp was identified encoding a predicted protein of 907 amino acids and a molecular mass of 104 kDa. A single transcript of approximately 3.2 kb was detected by Northern blot analysis. Comparison of the deduced amino acid sequence of the C. albicans NTC1 gene product with that of the Saccharomyces cerevisiae NTH1 gene product revealed 57% identity. The NTC1 gene was localized on chromosome 1 or R. A null mutant (Delta ntc1/Delta ntc1) was constructed by sequential gene disruption. Extracts from mutants homozygous for neutral trehalase deletion had only marginal neutral trehalase activity. Extracts from heterozygous mutants showed intermediate activities between extracts from the wild-type strain and from the homozygous mutants. The null mutant showed no significant differences in pathogenicity as compared to the wildtype strain in a mouse model of systemic candidiasis. This result indicates that the neutral trehalase of C. albicans is not a potential target for antifungal drugs.