The cell wall and cell division gene cluster in the Mra operon of Pseudomonas aeruginosa: cloning, production, and purification of active enzymes.
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.
Azzolina
B A
BA
Department of Biochemistry, Merck Research Laboratories, Rahway, New Jersey 07065, USA.
Yuan
X
X
Anderson
M S
MS
El-Sherbeini
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002455:Cell Division / Q000235:genetics; D002473:Cell Wall / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D021901:DNA, Intergenic / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D023061:Gene Order / Q000235:genetics; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics*; D010453:Peptide Synthases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000201:enzymology* / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid
D021901:DNA, Intergenic; D011994:Recombinant Proteins; D010453:Peptide Synthases