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利用lacZ融合技术来测定大肠杆菌unc操纵子前三个基因的体内表达情况。

Use of lacZ fusions to measure in vivo expression of the first three genes of the Escherichia coli unc operon.

作者信息

Solomon K A, Hsu D K, Brusilow W S

机构信息

Department of Chemistry and Biochemistry, University of Maryland, College Park 20742.

出版信息

J Bacteriol. 1989 Jun;171(6):3039-45. doi: 10.1128/jb.171.6.3039-3045.1989.

Abstract

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.

摘要

我们构建了与大肠杆菌unc操纵子的前三个基因的读框内lacZ蛋白融合体,该操纵子编码质子转运ATP酶的亚基。我们利用这些构建体来测量这些基因在体内的相对表达水平。编码F0区段的a和c亚基的第二个和第三个基因uncB和uncE,其相对表达水平约为1:10,不过uncB的测量表达水平取决于与lacZ融合的基因片段长度。这些表达率与纯化的F1F0复合物中a和c亚基的相对数量(a1:c10)相比情况良好。操纵子的第一个基因uncI在体内的表达非常低,最多比uncB的表达低10到20倍。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee0/210012/11b8345b73e7/jbacter00172-0143-a.jpg

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