Gunsalus R P, Brusilow W S, Simoni R D
Proc Natl Acad Sci U S A. 1982 Jan;79(2):320-4. doi: 10.1073/pnas.79.2.320.
We have constructed an extensive set of plasmids that carry the genes specifying the eight polypeptides of the proton-translocating ATPase of Escherichia coli. Using detailed restriction analysis and in vitro protein synthesis directed by these plasmids, we have established the order of the eight unc genes to be BEFHAGDC and the corresponding polypeptides to be a, c, b, delta, alpha, gamma, beta, and epsilon. These analyses include determining the location of the gene coding for the delta subunit of the F1 portion of the complex. We call this gene uncH. We have now established the gene order and gene-polypeptide relationships of the unc operon. This approach should be of use for study of other multigene bacterial operons, especially those with genes coding for polypeptides with unknown or unmeasurable catalytic activity.
我们构建了一系列广泛的质粒,这些质粒携带指定大肠杆菌质子转运ATP酶八种多肽的基因。通过详细的限制性分析以及由这些质粒指导的体外蛋白质合成,我们确定了八个unc基因的顺序为BEFHAGDC,相应的多肽顺序为a、c、b、δ、α、γ、β和ε。这些分析包括确定编码该复合物F1部分δ亚基的基因的位置。我们将这个基因称为uncH。我们现已确定了unc操纵子中的基因顺序和基因与多肽的关系。这种方法应用于研究其他多基因细菌操纵子,特别是那些编码具有未知或不可测量催化活性多肽的基因。