Wallace L J, Wilcox G
Mol Gen Genet. 1979 Jun 20;173(3):323-31. doi: 10.1007/BF00268643.
Hybrid plasmids were constructed from fragments of F'ara episomes formed by the restriction endonuclease EcoRI and a linear form of the plasmid ColE1 created by cleavage with EcoRI. Hybrid plasmids were constructed containing the entire ara region or the ara region with various parts deleted. E. coli K12 host strains were constructed which contained different deletions of the ara region. The hybrid plasmids were transferred to those strains whose ara deletion complemented that of the plasmid. The initial differential rates of synthesis of L-arabinose isomerase, the product of the araA gene, were determined for the Ara+, plasmid containing strains. These studies demonstrated that strains containing delta(araOIBA)718 produce elevated levels of araC protein, suggesting the araC promoter has been altered by this deletion. Evidence is also presented which suggests that araC protein activates the ara-BAD operon to higher levels when it is present in cis rather than trans. Amplification of the products of the cloned genes is observed when compared to haploid levels in some cases.
杂种质粒是由限制性内切酶EcoRI切割形成的F'ara附加体片段与用EcoRI切割产生的质粒ColE1线性形式构建而成。构建了包含整个ara区域或缺失了不同部分的ara区域的杂种质粒。构建了含有ara区域不同缺失的大肠杆菌K12宿主菌株。将杂种质粒转移到那些ara缺失与质粒互补的菌株中。对于含有Ara +质粒的菌株,测定了araA基因产物L-阿拉伯糖异构酶的初始合成差异速率。这些研究表明,含有delta(araOIBA)718的菌株产生的araC蛋白水平升高,表明该缺失改变了araC启动子。还提供了证据表明,araC蛋白以顺式而非反式存在时能将ara - BAD操纵子激活到更高水平。在某些情况下,与单倍体水平相比,观察到克隆基因产物的扩增。