Isolation and characterization of two chitin synthase genes from Aspergillus nidulans.
Isolation and characterization of two chitin synthase genes from Aspergillus nidulans.
复制标题
构巢曲霉中两个几丁质合酶基因的分离和表征。
DOI：
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
Masamichi Takagi
中科院分区：
文献类型：
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作者：
Koji Yanai;N. Kojima;Naoki Takaya;H. Horiuchi;A. Ohta;Masamichi Takagi
使用酿酒酵母 CHS2 基因作为杂交探针，从构巢曲霉中分离出两个几丁质合酶基因，命名为 chsA 和 chsB。核苷酸测序显示chsA和chsB分别编码由1013和916个氨基酸残基组成的多肽；这些酶的亲水特性与其他真菌几丁质合酶相似。 Northern 分析表明这两个基因均被转录，这表明构巢曲霉中的细胞几丁质是由至少两种几丁质合酶合成的。为了检查几丁质合酶基因在细胞生长中的作用，进行了基因破坏实验。 chsA 破坏株与野生型菌株一样生长，但 chsB 破坏株具有严重的生长缺陷，无法通过添加 1.2M 山梨醇作为渗透稳定剂来克服。这些发现表明 chsB 而不是 chsA 对于菌丝生长至关重要。
Two chitin synthase genes, designated chsA and chsB, were isolated from Aspergillus nidulans with the Saccharomyces cerevisiae CHS2 gene as the hybridization probe. Nucleotide sequencing showed that chsA and chsB encoded polypeptides consisting of 1013 and 916 amino acid residues, respectively; the hydropathy profiles of the enzymes were similar to those of other fungal chitin synthases. Northern analysis indicated that both genes were transcribed, suggesting that cellular chitin in A. nidulans is synthesized by at least two chitin synthases. For examination of the roles of the chitin synthase genes in cell growth, gene disruption experiments were done. The chsA disruptant grew as well as the wild-type strain, but the chsB disruptant had severe growth defects that could not be overcome by the addition of 1.2M sorbitol as an osmotic stabilizer. These findings suggested that chsB but not chsA is essential for hyphal growth.