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Nucleic Acids Research, 1984, Vol. 12, No. 15 6337-6355
© 1984

MOLECULAR BIOLOGY

Analysis of the Escherichia coli proBA locus by DNA and protein sequencing

A.H. Deutch1,*, K.E. Rushlow2 and C.J. Smith3

Bethesda Research Laboratories, Inc. P.O. Box 6009, Gaithersburg, MD 20877, USA

*To whom correspondence sould be addressed

Received March 6, 1984. Revised July 10, 1984. Accepted July 10, 1984.

A 2.9 kb DNA fragment carrying the Escherichia coli proBA region, which encodes the first two enzymes of the proline biosynthetic pathway, was subcloned onto an expression plasmid carrying both the bacteriophage lambda PLpromoter ({lambda}PL) and the lambda gene encoding a thermolabile cI repressor protein (cI857). Derepression of the {lambda}PL promoter by thermal inactivation of the cI857 repressor protein resulted in the simultaneous overproduction of the proB ({gamma}-glutamyl kinase) and proA ({gamma}-glutamyl phosphate reductase) gene products. Nucleotide sequence analysis of the proBA locus allowed gene assignments consistent with the NH2 and COOH-terminal analyses and amino acid compositions of homogeneous preparations of the proB and proA proteins. The contiguous nature of the proB and proA genes suggests that the two genes constitute an operon in which proB precedes proA.

1Present address: Washington Research Center, W.R.Grace & Co., 7379 Route 32, Columbia, MD 21044, USA

2Present address: Syngene Products & Research, 225 Commerce Drive, P.O. Box 2211, Fort Collins, CO 80522, USA

3Present address: Laboratory of Molecular Microbiology, NIAID, Building 550, Fort Detrick, Frederick, MD 21701, USA


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