Plasmid vectors for the selection of promoters.

Vectors were constructed which contain promoterless genes for chloramphenicol (cam) or tetracycline (tet) resistance, as promoter-probe plasmids. Escherichia coli cells harboring these plasmids are sensitive to cam or tet but resistant to ampicillin. In plasmids pKK231 -1 and pKK232 -8 the gene for cam acetyltransferase (CAT) and in pKK175 -6 the gene for tet resistance are flanked by efficient transcription terminators, preventing transcription from other pBR322 promoters into the antibiotic resistance region. In one of the vectors, pKK232 -8, translational stop codons were introduced in all three reading frames upstream from the initiation codon of the cat gene. If a DNA fragment containing a promoter is inserted into one of the cloning sites upstream from the antibiotic genes, cells carrying such plasmids acquire resistance to cam or tet. Using these vectors two restriction fragments that contain promoters were identified. One of these fragments contains sequences upstream from an unidentified gene ( ORFII ) located distal to the rrnB rRNA operon of E. coli.

[1]  R. Neve,et al.  Construction and characterization of E. coli promoter-probe plasmid vectors. I. Cloning of promoter-containing DNA fragments. , 1979, Gene.

[2]  R. Rodriguez,et al.  Construction and characterization of E. coli promoter-probe plasmid vectors. III. pBR322 derivatives with deletions in the tetracycline resistance promoter region. , 1982, Gene.

[3]  G. Cesareni,et al.  Control of ColE1 DNA replication: the rop gene product negatively affects transcription from the replication primer promoter. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[4]  W. Gilbert,et al.  Eukaryotic signal sequence transports insulin antigen in Escherichia coli. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. Vieira,et al.  A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. , 1982, Gene.

[6]  P. Sharp,et al.  Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids , 1977, Cell.

[7]  H. Birnboim,et al.  A rapid alkaline extraction procedure for screening recombinant plasmid DNA. , 1979, Nucleic acids research.

[8]  W. Gilbert,et al.  An ‘internal” signal sequence directs secretion and processing of proinsulin in bacteria , 1981, Nature.

[9]  D. Sherratt,et al.  Trans-complementable copy-number mutants of plasmid ColE1 , 1980, Nature.

[10]  L. Comstock,et al.  The tac promoter: a functional hybrid derived from the trp and lac promoters. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[11]  H. Bujard,et al.  Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid specified genes. , 1982, The EMBO journal.

[12]  H. Bujard,et al.  Cloning and analysis of strong promoters is made possible by the downstream placement of a RNA termination signal. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Legerski,et al.  Extracellular nucleases of Pseudomonas BAL 31. I. Characterization of single strand-specific deoxyriboendonuclease and double-strand deoxyriboexonuclease activities. , 1975, Nucleic acids research.

[14]  W. Fiers,et al.  Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. , 1981, Gene.

[15]  S. Falkow,et al.  Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. , 1977, Gene.

[16]  R. Rodriguez,et al.  Eukaryotic DNA fragments which act as promoters for a plasmid gene , 1979, Nature.

[17]  H. Noller,et al.  Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. , 1981, Journal of molecular biology.

[18]  H. Schaller,et al.  Segment-specific mutagenesis: extensive mutagenesis of a lac promoter/operator element. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[19]  D. Pribnow.  Genetic Control Signals in DNA , 1979 .

[20]  Walter Gilbert,et al.  E. coli RNA polymerase interacts homologously with two different promoters , 1980, Cell.

[21]  G. An,et al.  Plasmid vehicles for direct cloning of Escherichia coli promoters , 1979, Journal of bacteriology.

[22]  R. Gutell,et al.  Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. , 1981, Plasmid.

[23]  J. Brosius.  Toxicity of an overproduced foreign gene product in Escherichia coli and its use in plasmid vectors for the selection of transcription terminators. , 1984, Gene.

[24]  D Court,et al.  Regulatory sequences involved in the promotion and termination of RNA transcription. , 1979, Annual review of genetics.

[25]  F. Bolivar,et al.  Characterizing wild-type and mutant promoters of the tetracycline resistance gene in pBR313. , 1979, Nucleic acids research.

[26]  R. Gutell,et al.  Transcriptional mapping of plasmid pKK3535. Quantitation of the effect of guanosine tetraphosphate on binding to the rrnB promoters and a lambda promoter with sequence homologies in the CII binding region. , 1981, Journal of molecular biology.

[27]  D. Vapnek,et al.  Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9 , 1979, Nature.

[28]  W. Gilbert,et al.  Sequencing end-labeled DNA with base-specific chemical cleavages. , 1980, Methods in enzymology.

[29]  J. Sutcliffe.  Complete nucleotide sequence of the Escherichia coli plasmid pBR322. , 1979, Cold Spring Harbor symposia on quantitative biology.

[30]  D. Goldfarb,et al.  Expression of Tn9-derived chloramphenicol resistance in Bacillus subtilis , 1981, Nature.

[31]  J. Brosius,et al.  Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. , 1983, Gene.

[32]  S. Colowick,et al.  Methods in Enzymology , Vol , 1966 .

[33]  Stanley N Cohen,et al.  Phenotypic expression in E. coli of a DNA sequence coding for mouse dihydrofolate reductase , 1978, Nature.

[34]  J. Brosius,et al.  Precise location of two promoters for the beta-lactamase gene of pBR322. S1 mapping of ribonucleic acid isolated from Escherichia coli or synthesized in vitro. , 1982, The Journal of biological chemistry.

[35]  D. Schümperli,et al.  Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon , 1982, Cell.

[36]  K. Talmadge,et al.  Construction of plasmid vectors with unique PstI cloning sites in a signal sequence coding region. , 1980, Gene.

[37]  T. Close,et al.  Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. , 1982, Gene.

[38]  H. Bujard,et al.  Organization of transcriptional signals in plasmids pBR322 and pACYC184. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[39]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.