Abstract
The 2-min region of the Escherichia coli genome contains a large cluster of genes from pbpB to envA that code for proteins involved in peptidoglycan biosynthesis and cell division. From pLC26-6 of the collection of Clarke and Carbon (L. Clarke and J. Carbon, Cell 9:91-99, 1976) plasmids carrying different fragments from the 8-kilobase-pair region downstream of pbpB were constructed and analyzed for their ability to direct protein synthesis in maxicells, to complement various thermosensitive mutations, and to overproduce enzymatic activities. We report the localization of the previously unidentified murD gene coding for the D-glutamic acid-adding enzyme within this region. Our data show that the genes are in the order pbpB-murE-murF-X-murD-Y-murG, where X and Y represent chromosomal fragments from 1 to 1.5 kilobase pairs, possibly coding for unknown proteins. Furthermore, the murE and murF genes, encoding the meso-diaminopimelic acid and D-alanyl-D-alanine-adding enzymes, respectively, may be translationally coupled when transcription is initiated upstream of murE, within the preceding structural gene pbpB coding for penicillin-binding protein 3.