Regular Article
The Cell Wall and Cell Division Gene Cluster in the Mra Operon of Pseudomonas aeruginosa: Cloning, Production, and Purification of Active Enzymes

https://doi.org/10.1006/prep.2001.1390Get rights and content

Abstract

We have cloned the Pseudomonas aeruginosa cell wall biosynthesis and cell division gene cluster that corresponds to the mra operon in the 2-min region of the Escherichia coli chromosome. The organization of the two chromosomal regions in P. aeruginosa and E. coli is remarkably similar with the following gene order: pbp3/pbpB, murE, murF, mraY, murD, ftsW, murG, murC, ddlB, ftsQ, ftsA, ftsZ, and envA/LpxC. All of the above P. aeruginosa genes are transcribed from the same strand of DNA with very small, if any, intragenic regions, indicating that these genes may constitute a single operon. All five amino acid ligases, MurC, MurD, MurE, MurF, and DdlB, in addition to MurG and MraY were cloned in expression vectors. The four recombinant P. aeruginosa Mur ligases, MurC, MurD, MurE, and MurF were overproduced in E. coli and purified as active enzymes.

References (28)

  • C. Parquet et al.

    Nucleotide sequence of the murF gene encoding the UDP-MurNAc-pentapeptide synthetase of Escherichia coli.

    Nucleic Acids Res.

    (1989)
  • A.C. Robinson et al.

    Further evidence for overlapping transcriptional units in an Escherichia coli cell envelope-cell division gene cluster: DNA sequence and transcriptional organization of the ddl ftsQ region

    J. Bacteriol.

    (1986)
  • M. Ikeda et al.

    The Escherichia coli mraY gene encoding UDP-N-acetylmuramoyl-pentapeptide:Undecaprenyl-phosphatephospho-N-acetylmuramoyl-pentapeptide transferase

    J. Bacteriol.

    (1991)
  • Cited by (0)

    1

    To whom correspondence and reprint reqquests should be addressed. Fax: (732) 594-5878.

    View full text