Miyashita K, Fujii T
National Institute of Agro-Environmental Sciences, Ibaraki, Japan.
Biosci Biotechnol Biochem. 1993 Oct;57(10):1691-8. doi: 10.1271/bbb.57.1691.
A chitinase gene (chiA) from Streptomyces lividans was characterized and its nucleotides sequenced. Although the deduced amino acid sequence of chitinase A1 did not show any similarity to those of other Streptomyces chitinases that has been sequenced, the C-terminal part, containing both a putative catalytic domain and type-III-like repeating units, showed a similarity (36%) to that of chitinase D from Bacillus circulans. A site of initiation of transcription was found approximately 51 bp upstream from the GTG initiation codon. The promoter region of the chiA gene was subcloned on a 178-bp fragment into the promoter-probe vector pIJ486, resulting in the chitin stimulated expression of the neomycin resistance gene. One of the deleted subclones, which contained a 114-bp sequence upstream from the translation start codon, retained both chitin stimulated production and glucose repression. Chitin stimulated production was lost in an other deleted mutant containing the 104-bp upstream sequence.
对来自浅青紫链霉菌的几丁质酶基因（chiA）进行了表征并测定了其核苷酸序列。尽管几丁质酶A1推导的氨基酸序列与已测序的其他链霉菌几丁质酶的氨基酸序列没有任何相似性，但包含推定催化结构域和III型样重复单元的C末端部分与环状芽孢杆菌的几丁质酶D的C末端部分显示出相似性（36%）。在GTG起始密码子上游约51 bp处发现了一个转录起始位点。chiA基因的启动子区域被亚克隆到一个178 bp的片段上，插入到启动子探针载体pIJ486中，导致几丁质刺激新霉素抗性基因的表达。其中一个缺失亚克隆包含翻译起始密码子上游114 bp的序列，保留了几丁质刺激产生和葡萄糖阻遏的特性。几丁质刺激产生在另一个包含104 bp上游序列的缺失突变体中消失。
