Function of Corynebacterium glutamicum promoters in Escherichia coli, Streptomyces lividans, and Bacillus subtilis.

The function of seven promoters from Corynebacterium glutamicum, P-hom, P-leuA, P-per, P-aes1, P-aes2, P-45, and P-104, was analyzed in a heterologous background. DNA fragments carrying the promoters were cloned into shuttle promoter-probe vectors replicating in Escherichia coli and C. glutamicum (pET2), Streptomyces lividans (pGL7011) and Bacillus subtilis (pRB394). With the exception of P-hom, P-leuA and P-104 in B. subtilis, all promoters were found to be active in all species. Non-radioactive methods of primer-extension analysis and of S1-nuclease protection assay using automatic sequencer were developed to determine the respective transcriptional start points (TSPs). All TSPs were determined by primer extension and in two promoters (P-45 and P-hom) the main TSPs were confirmed by S1-mapping. While the main TSPs were identical in all four species, utilization of multiple TSPs varied among the species and additional TSPs were detected in S. lividans. Knowledge of the efficiency of promoters and of exact respective TSPs may be of practical value for the construction of expression systems in a heterologous background.

[1]  R. Brückner,et al.  A series of shuttle vectors for Bacillus subtilis and Escherichia coli. , 1992, Gene.

[2]  J. Vohradský,et al.  Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number , 1997, Journal of bacteriology.

[3]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[4]  M. Wösten.  Eubacterial sigma-factors. , 1998, FEMS microbiology reviews.

[5]  M. Bibb,et al.  Isolation and characterization of a strong promoter element from the Streptomyces ghanaensis phage I19 using the gentamicin resistance gene (aacC1) of Tn 1696 as reporter. , 1997, Microbiology.

[6]  M. Voskuil,et al.  Rapid isolation and sequencing of purified plasmid DNA from Bacillus subtilis , 1993, Applied and environmental microbiology.

[7]  J. Trevors,et al.  Bacterial conjugation between pseudomonads in the rhizosphere of wheat , 1988 .

[8]  Akira Ishihama,et al.  Micrococcus luteus, a bacterium with a high genomic G+C content, contains Escherichia coli-type promoters , 1989, Molecular and General Genetics MGG.

[9]  G. Frey,et al.  Control regions of an archaeal gene. A TATA box and an initiator element promote cell-free transcription of the tRNA(Val) gene of Methanococcus vannielii. , 1991, Journal of molecular biology.

[10]  H. Sahm,et al.  Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. , 1994, Microbiology.

[11]  R. Doi,et al.  Prokaryotic promoters in biotechnology. , 1995, Biotechnology annual review.

[12]  W R Strohl,et al.  Compilation and analysis of DNA sequences associated with apparent streptomycete promoters. , 1992, Nucleic acids research.

[13]  M. Pátek,et al.  Integrative and autonomously replicating vectors for analysis of promoters in Corynebacterium glutamicum , 1998 .

[14]  M. Bashyam,et al.  A study of mycobacterial transcriptional apparatus: identification of novel features in promoter elements , 1996, Journal of bacteriology.

[15]  M. Malumbres,et al.  Transcriptional analysis and regulatory signals of the hom-thrB cluster of Brevibacterium lactofermentum , 1994, Journal of bacteriology.

[16]  J. Bailey,et al.  Toward a science of metabolic engineering , 1991, Science.

[17]  M. Pátek,et al.  Leucine synthesis in Corynebacterium glutamicum: enzyme activities, structure of leuA, and effect of leuA inactivation on lysine synthesis , 1994, Applied and environmental microbiology.

[18]  R. Katsumata,et al.  Functional expression of the genes of Escherichia coli in gram-positive Corynebacterium glutamicum , 2004, Molecular and General Genetics MGG.

[19]  M. Pátek,et al.  Identification of a novel gene involved in stable maintenance of plasmid pGA1 from Corynebacterium glutamicum. , 2001, Plasmid.

[20]  M. Pátek,et al.  Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif. , 1996, Microbiology.

[21]  M. Bibb,et al.  Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. , 1983, Journal of general microbiology.

[22]  M. Pátek,et al.  Promoters of Corynebacterium glutamicum. , 2003, Journal of biotechnology.

[23]  H. Margalit,et al.  Determination of common structural features in Escherichia coli promoters by computer analysis. , 1994, European journal of biochemistry.

[24]  M. Pátek,et al.  Analysis of the Corynebacterium glutamicum dapA Promoter , 1999, Journal of bacteriology.

[25]  B. Jaurin,et al.  Streptococcus pneumoniae possesses canonical Escherichia coli (sigma 70) promoters , 1990, Molecular microbiology.

[26]  A. Pühler,et al.  The Corynebacterium glutamicum insertion sequence ISCg2 prefers conserved target sequences located adjacent to genes involved in aspartate and glutamate metabolism , 1999, Molecular and General Genetics MGG.

[27]  H. Margalit,et al.  Compilation of E. coli mRNA promoter sequences. , 1993, Nucleic acids research.

[28]  J. D. Helmann,et al.  Compilation and analysis of Bacillus subtilis sigma A-dependent promoter sequences: evidence for extended contact between RNA polymerase and upstream promoter DNA , 1995, Nucleic Acids Res..

[29]  H. Sahm,et al.  Analysis of a Corynebacterium glutamicum hom gene coding for a feedback-resistant homoserine dehydrogenase , 1991, Journal of bacteriology.

[30]  H Sahm,et al.  the Czech Republic, , 2022 .

[31]  D. Hopwood.  Genetic manipulation of Streptomyces : a laboratory manual , 1985 .

[32]  K. Schleifer,et al.  High efficiency electroporation of intact Corynebacterium glutamicum cells. , 1989, FEMS microbiology letters.

[33]  William R. Bourn,et al.  Computer assisted identification and classification of streptomycete promoters. , 1995, Nucleic acids research.

[34]  Stanley N Cohen,et al.  High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA , 1979, Molecular and General Genetics MGG.

[35]  E. Grohmann,et al.  Determination of specific DNA strand discontinuities with nucleotide resolution in exponentionally growing bacteria harboring rolling circle-replicating plasmids. , 1997, FEMS microbiology letters.