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白色念珠菌几丁质合成酶3基因的克隆及其在酵母-菌丝转变过程中的表达。

Cloning of the chitin synthase 3 gene from Candida albicans and its expression during yeast-hyphal transition.

作者信息

Sudoh M, Nagahashi S, Doi M, Ohta A, Takagi M, Arisawa M

机构信息

Department of Mycology, Nippon Roche Research Center, Kanagawa, Japan.

出版信息

Mol Gen Genet. 1993 Nov;241(3-4):351-8. doi: 10.1007/BF00284688.

Abstract

The chitin synthase 3 gene (CACHS3) has been cloned from Candida albicans. The yeast CAL1 gene encoding the chitin synthase 3 of Saccharomyces cerevisiae was used as a probe for the isolation of the gene from C. albicans. The CAL1 homolog was identified in Southern blots of C. albicans genomic DNA and cloned from a C. albicans genomic DNA library. The nucleotide sequences of two partial clones were determined and combined giving a total length of 4610 bp. A continuous open reading frame of 3525 bp encoding a predicted protein of 1175 amino acids and molecular mass of 131 850 daltons was identified. A comparison of the deduced amino acid sequences of CAL1 and the Candida chitin synthase 3 protein showed 59.3% identity. Southern blot analysis indicates that the CACHS3 gene is present in a single copy in the genome and maps to chromosome I. Northern blot analysis shows that expression of chitin synthase 3 gene is dramatically increased during the transition from the yeast to the hyphal form of C. albicans. This change in transcription level strongly suggests that CACHS3 may play a role in Candida morphogenesis.

摘要

几丁质合成酶3基因(CACHS3)已从白色念珠菌中克隆出来。酿酒酵母中编码几丁质合成酶3的酵母CAL1基因被用作从白色念珠菌中分离该基因的探针。在白色念珠菌基因组DNA的Southern印迹中鉴定出CAL1同源物,并从白色念珠菌基因组DNA文库中克隆出来。测定并合并了两个部分克隆的核苷酸序列,总长度为4610 bp。鉴定出一个3525 bp的连续开放阅读框,编码一个预测的1175个氨基酸的蛋白质,分子量为131850道尔顿。CAL1推导的氨基酸序列与白色念珠菌几丁质合成酶3蛋白的比较显示出59.3%的同一性。Southern印迹分析表明,CACHS3基因在基因组中以单拷贝形式存在,并定位于染色体I。Northern印迹分析表明,在白色念珠菌从酵母形态转变为菌丝形态的过程中,几丁质合成酶3基因的表达显著增加。转录水平的这种变化强烈表明,CACHS3可能在念珠菌形态发生中起作用。

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