CHARACTERIZATION AND EXPRESSION OF A P-450-LIKE MYCINAMICIN BIOSYNTHESIS GENE USING A NOVEL MICROMONOSPORA ESCHERICHIA-COLI SHUTTLE COSMID VECTOR
CHARACTERIZATION AND EXPRESSION OF A P-450-LIKE MYCINAMICIN BIOSYNTHESIS GENE USING A NOVEL MICROMONOSPORA ESCHERICHIA-COLI SHUTTLE COSMID VECTOR
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DOI：
10.1007/bf00302258
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发表时间：
1994-11-15
期刊：
影响因子：
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通讯作者：
HORINOUCHI, S
中科院分区：
文献类型：
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作者：
INOUYE, M;TAKADA, Y;HORINOUCHI, S
从隐藏的灰红小单孢菌质粒和大肠杆菌粘粒pJB 8构建了29 kb穿梭粘粒载体pTYS 507。在pTYS 507中亚克隆麦司霉素II生物合成基因，鉴定出一个可与M突变体互补的DNA区域。griseorubida缺乏羟化酶和环氧酶活性。核苷酸序列和突变分析表明，单个P-450样蛋白催化这两种反应。
A 29 kb shuttle cosmid vector, pTYS507, was constructed from a cryptic Micromonospora griseorubida plasmid and the Escherichia coli cosmid pJB8. Subcloning of mycinamicin II biosysnthesis genes in pTYS507 led to the identification of a DNA region that could complement a mutant of M. griseorubida that lacked both hydroxylase and epoxidase activities. Nucleotide sequence and mutational analysis suggested that a single P-450-like protein catalyzes both reactions.