MOLECULAR-CLONING AND CHARACTERIZATION OF THE ALKB GENE OF ESCHERICHIA-COLI
MOLECULAR-CLONING AND CHARACTERIZATION OF THE ALKB GENE OF ESCHERICHIA-COLI
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DOI：
10.1007/bf00383004
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发表时间：
1985-01-01
期刊：
影响因子：
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通讯作者：
SEKIGUCHI, M
中科院分区：
文献类型：
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作者：
KATAOKA, H;SEKIGUCHI, M
利用体外重组的方法，构建了能够抑制alkB突变引起的甲基磺酸敏感性增加的杂合质粒。由于克隆的DNA片段在47分钟时被定位在大肠杆菌K12遗传图谱上，即alkB基因所在的区域，因此可以得出结论，克隆的DNA片段包含alkB基因本身，但不包含抑制alkB突变的其他基因。通过 maxicell 方法对质粒编码的蛋白质进行特异性标记表明 alkB 编码分子量为 .apprx 的多肽。 27,000。在细菌染色体的 alkB 区域引入一个小缺失会导致 alkB 和 ada 基因失活，表明这两个基因在大肠杆菌染色体上相邻。
Using methods of an vitro recombination, one constructed hybrid plasmids that can suppress the increased methylmethane sulfonate sensitivity caused by alkB mutation. Since the cloned DNA fragment was mapped at 47 min on the E. coli K12 genetic map, an area where the alkB gene is located, it was concluded that the cloned DNA fragment contains the alkB gene itself, but not other genes that suppress alkB mutation. Specific labeling of plasmid-encoded proteins by the maxicell method revealed that the alkB codes for a polypeptide with a MW of .apprx. 27,000. Introduction of a small deletion into the alkB region of the bacterial chromosome resulted in inactivation of the alkB and ada genes, suggesting that the 2 genes are adjacent on the E. coli chromosome.