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A C Porter

Publications and source records attributed to A C Porter.

39 records · Page 3Linked to original sources

Cloning and expression of uncI, the first gene of the unc operon of Escherichia coli.

The unc operon of Escherichia coli consists of eight genes coding for the eight subunits of the proton-translocating ATPase. In vitro transcription-translation of DNA cloned from the beginning of the operon onto plasmids reveals that the reading frame uncI, which precedes the other genes of the operon, codes for a protein with a molecular weight of 14,500, called i. In minicells, the i protein is synthesized in amounts comparable to the amounts of the ATPase subunits, suggesting that it may be part of the ATPase complex. The presence of the unc promoter and uncI on a plasmid containing the other eight genes of the unc operon has little effect on the differential expression of the unc genes or the partitioning of the newly synthesized subunits into soluble or sedimentable fractions in the in vitro system. The i protein partitions into the sedimentable fraction.

Adenosine Triphosphatases↗

Promoter for the unc operon of Escherichia coli.

Fragments of DNA carrying possible promoters for the unc operon of Escherichia coli were cloned into a promoter detection plasmid (pRZ5255). Similar fragments were transcribed in vitro to produce transcripts whose sizes were used to determine the approximate start site for transcription. One strong promoter and at least two very much weaker ones were detected by these methods. The exact position of the strongest promoter, presumed to be the true unc promoter, was determined by S1 nuclease mapping and shown to lie 73 base pairs upstream from the open reading frame that precedes uncB. It therefore appears that this reading frame (uncI) is part of the unc operon. S1 mapping also revealed the presence of a third weak promoter 25 base pairs upstream of uncI. All of the weak promoters occur between the proposed unc promoter and uncB, but their role in vivo, if any, is unclear.

Adenosine Triphosphatases↗