Abstract
The primary goal of our work is to contribute to a better understanding of structure-function relationships of nucleotide sequences defining promoters in theE. coli system. Since strong, unregulated promoters can be assumed to exhibit pertinent structural features most clearly, we investigated, in vitro, more than 50 promoters for their ability (a) to compete for the binding of RNA polymerase (an indication of their “signal strength”) and (b) to produce RNA (“overall efficiency” of a promoter) under conditions of limiting amounts of RNA polymerase. Promoters fulfilling these requirements best were found primarily in the genome of coliphage T5 (1). Some of these promoters show extremely high rates of complex formation with RNA polymerase (1,2) and at the same time were identified as being very efficient initiators of RNA synthesis in vitro and in vivo (1,3).
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References
Gabain, A.V., and H. Bujard (1979) Interaction ofE.coli RNA polymerase with promoters: The relative rates of complex formation and decay with promoters of several phage and plasmid DNA’s.Proc. Natl. Acad. Sci., USA 76:189–193.
Brunner, M. (1984) Untersuchungen zur Wechselwirkung vonE.coli RNA-Polymerase mit Promotoren. Diplomarbeit, Universität Heidelberg, 69 Heidelberg, FRG.
Gentz, R., A. Langner, A.C.Y. Chang, S.N. Cohen, and H. Bujard (1981) Cloning and analysis of strong promoters is made possible by the downstream placement of an RNA termination signal.Proc. Natl. Acad. Sci., USA 78:4936–4941.
Stueber, D., and H. Bujard (1982) Transcription from efficient promoters can interfere with plasmid replication and diminish expression of plasmid specified genes.EMBO 1:1399–1404.
Cesarini, G., M.A. Muesing, and B. Polisky (1982) Control of ColE1 DNA replication: The Rop gene product negatively affects transcription from the replication primer promoter.Proc. Natl. Acad. Sci., USA 79:6313–6317.
Muesing, M., J. Tamm, H.M. Shepard, and B. Polisky (1981) A single base-pair alternation is responsible for the DNA overproduction phenotype of a plasmid copy-number mutant.Cell 24:235–242.
Tomizawa, J., and T. Itoh (1981) Plasmid ColE1 incompatibility determined by interaction of RNA I with primer transcript.Proc. Natl. Acad. Sci., USA 78:6096–6100.
Stueber, D., and H. Bujard (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184.Proc. Natl. Acad. Sci., USA 78:167–171.
Deuschle, U. (1984) Entwicklung einer Methode zur quantitativen Bestimmung der in vitro— als auchin vivo-Effizienzen von Transkriptionssignalen aus demE.coli-system.Diplomarbeit, Universität Heidelberg, 69 Heidelberg, FRG.
Deuschle, U., R. Gentz, and H. Bujard (unpublished results).
Summers, D.K., and D.J. Sherrat (1984) Multimerization of high copy number plasmids causes instability: ColE1 encodes a determinant essential for plasmid monomerization and stability. Cell 36:1097–1103.
Peschke, U. (1984) Konstruction von tandemartig angeordneten Promoter/Operator sequenzen zur analyse der wechselwirkung diser sequenzen mit RNA-polymerase bzw. Repressoren.Diplomarbeit, Universität Heidelberg, 69 Heidelberg, FRG.
Bujard, H., C. Baldari, M. Brunner, U. Deuschle, R. Gentz, J. Hughes, W. Kammerer, and D. Stueber (1983) Integration of efficient promoters of the E. coli system into plasmid vectors. InGene Amplification and Analysis, T.S. Papas, M. Rosenberg, and J.C. Cirikjian, eds. Elsevier/North Holland, New York, Vol. 3, pp. 65–88.
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© 1985 Plenum Press, New York
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Bujard, H., Stueber, D., Gentz, R., Deuschle, U., Peschke, U. (1985). Insertion of Transcriptional Elements Outside the Replication Region can Interfere with Replication, Maintenance, and Stability of ColE1-Derived Plasmids. In: Helinski, D.R., Cohen, S.N., Clewell, D.B., Jackson, D.A., Hollaender, A. (eds) Plasmids in Bacteria. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2447-8_6
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DOI: https://doi.org/10.1007/978-1-4613-2447-8_6
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