Deletion and mutational analysis of the promoter P-dapA from Corynebacterium glutamicum was performed to identify regions and particular nucleotides important for its function. An extended -10 region and a stretch of six T's at positions -55 to -50 were found to be the most important elements in the promoter function. The results of mutational analysis of P-dapA are consistent with the conclusions of statistical computer-aided analysis of 44 C. glutamicum promoter sequences.

Institute of Microbiology Academy of Sciences of the Czech Republic CZ 14220 Praha 4 Czech Republic

Zobrazit více v PubMed

Galas D J, Eggert M, Waterman M S. Rigorous pattern-recognition methods for DNA sequences. J Mol Biol. 1985;186:117–128. PubMed

Graves M C, Rabinowitz J C. In vivo and in vitro transcription of the Clostridium pasteurianum ferredoxin gene. Evidence for “extended” promoter elements in gram-positive organisms. J Biol Chem. 1986;261:11409–11415. PubMed

Harley C B, Reynolds R P. Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987;15:2343–2361. PubMed PMC

Helmann J D. Compilation and analysis of Bacillus subtilis sigma A-dependent promoter sequences: evidence for extended contact between RNA polymerase and upstream promoter DNA. Nucleic Acids Res. 1995;23:2351–2360. PubMed PMC

Ito W, Ishiguro H, Kurosawa Y. A general method for introducing a series of mutations into cloned DNA using the polymerase chain reaction. Gene. 1991;102:67–70. PubMed

Keilhauer C, Eggeling L, Sahm H. Isoleucine synthesis in Corynebacterium glutamicum: molecular analysis of the ilvB-ilvN-ilvC operon. J Bacteriol. 1993;175:5595–5603. PubMed PMC

Keilty S, Rosenberg M. Constitutive function of a positively regulated promoter reveals new sequences essential for activity. J Biol Chem. 1987;262:6389–6395. PubMed

Kumar A, Malloch R A, Fujita N, Smillie D A, Ishihama A, Hayward R S. The minus 35-recognition region of Escherichia coli sigma 70 is inessential for initiation of transcription at an “extended minus 10” promoter. J Mol Biol. 1993;232:406–418. PubMed

Nešvera J, Hochmannová J, Pátek M. An integron of class 1 is present on the plasmid pCG4 from Gram-positive bacterium Corynebacterium glutamicum. FEMS Microbiol Lett. 1998;169:391–395. PubMed

Pátek M, Eikmanns B J, Pátek J, Sahm H. Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif. Microbiology. 1996;142:1297–1309. PubMed

Pátek M, Nešvera J, Hochmannová J, Štokrová J. Transfection of Brevibacterium flavum with bacteriophage BFB10 DNA. Folia Microbiol. 1988;33:247–254.

Perez-Martin J, de Lorenzo V. Clues and consequences of DNA bending in transcription. Annu Rev Microbiol. 1997;51:593–628. PubMed

Ross W, Gosink K K, Salomon J, Igarashi K, Zou C, Ishihama A, Severinov K, Gourse R L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993;262:1407–1413. PubMed

Sahm H, Eggeling L, Eikmanns B, Krämer R. Metabolic design in amino acid-producing bacterium Corynebacterium glutamicum. FEMS Microbiol Rev. 1995;16:243–252.

Schäfer A, Kalinowski J, Simon R, Seep-Feldhaus A H, Pühler A. High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria. J Bacteriol. 1990;172:1663–1666. PubMed PMC

Shaw W V. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 1975;43:737–755. PubMed

Vašicová P, Abrhámová Z, Nešvera J, Pátek M, Sahm H, Eikmanns B. Integrative and autonomously replicating vectors for analysis of promoters in Corynebacterium glutamicum. Biotechnol Tech. 1998;12:743–746.

Corynebacterium glutamicum promoters: a practical approach

General and molecular microbiology and microbial genetics in the IM CAS

Plasmid vectors for testing in vivo promoter activities in Corynebacterium glutamicum and Rhodococcus erythropolis

Control of rep gene expression in plasmid pGA1 from Corynebacterium glutamicum