Dutka-Malen S, Mazodier P, Badet B
Laboratoire de Bioorganique & Biotechnologies, ENSCP, Paris, France.
Biochimie. 1988 Feb;70(2):287-90. doi: 10.1016/0300-9084(88)90073-9.
A recombinant plasmid carrying a 4.6 kg restriction endonuclease NcoI-ClaI fragment of genomic DNA from Escherichia coli K12 was constructed. This plasmid complements the glmS mutation. Subcloning into pUC18 gave plasmid pGM10 encoding the structural gene of glucosamine synthetase, as judged by overexpression of enzyme activity and the isolation in high yield of the pure protein.
构建了一个携带来自大肠杆菌K12基因组DNA的4.6 kb限制性内切酶NcoI-ClaI片段的重组质粒。该质粒可互补glmS突变。通过酶活性的过表达以及纯蛋白的高产率分离判断，亚克隆到pUC18中得到了编码氨基葡萄糖合成酶结构基因的质粒pGM10。
