Use of lacZ fusions to measure in vivo expression of the first three genes of the Escherichia coli unc operon.
We have constructed in-frame lacZ protein fusions to the first three genes of the Escherichia coli unc operon, which codes for the subunits of the proton-translocating ATPase. We have used these constructions to measure the relative in vivo expression of these genes. The second and third genes, uncB and uncE, which code for the a and c subunits of the F0 sector, were expressed at relative levels of approximately 1:10, although the measured expression of uncB depended upon how much of the gene was fused to lacZ. These rates compared favorably with the relative numbers of a and c subunits (a1:c10) in the purified F1F0 complex. The in vivo expression of uncI, the first gene of the operon, was very low, at best 10 to 20 times less than the expression of uncB.
Solomon
K A
KA
Department of Chemistry and Biochemistry, University of Maryland, College Park 20742.
Hsu
D K
DK
Brusilow
W S
WS
D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D006180:Proton-Translocating ATPases / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D006180:Proton-Translocating ATPases