Cloning and expression of the ilvB gene of Escherichia coli K-12.
Cloning and expression of the ilvB gene of Escherichia coli K-12.
复制标题
大肠杆菌K-12 ilvB 基因的克隆和表达。
DOI：
10.1007/bf00729457
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Freundlich,M
中科院分区：
文献类型：
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作者：
Newman,T;Friden,P;Sutton,A;Freundlich,M
用内切酶esali处理过的pBR322质粒和filvb4质粒的DNA连接混合物分离出了一个含有ilvboperon的质粒，该质粒编码大肠杆菌k -12的乙酰羟基酸合成酶I。将一个3.4 kb的细菌片段克隆到质粒pKEN005中，得到该质粒的缩短衍生物pTCN12。通过限制性内切酶定位确定了它们的ilvboperon相对于质粒基因的定位。对ilvbgene产物乙酰羟基酸合成酶I的水平测定表明，质粒pTCN12含有一个功能性的ilvb启动子和控制区。用该质粒的DNA作为探针，表明合成ofilvBmRNA的速率与乙酰羟基酸合酶I的水平成正比。
A plasmid containing theilvBoperon, which codes for acetohydroxy acid synthase I ofEscherichia coliK-12, was isolated using a ligated mixture of DNA from plasmid pBR322 and F'ilvB4treated with endonucleaseSalI. A shortened derivative of this plasmid was isolated by cloning a 3.4 kb bacterial fragment into plasmid pKEN005 to yield plasmid pTCN12. The orientation of theilvBoperon relative to plasmid genes was determined by restriction enzyme mapping. Measurement of the level of the product of theilvBgene, acetohydroxy acid synthase I, indicated that plasmid pTCN12 contained a functionalilvBpromoter and control region. The DNA from this plasmid was used as a probe to show that the rate of synthesis ofilvBmRNA was proportional to the levels of acetohydroxy acid synthase I.