Regions of broad-host-range plasmid RK2 which are essential for replication and maintenance
TLDR
The results indicate that at least two plasmid-encoded genes, capable of acting in trans, and a replication origin are required for RK2 replication and maintenance.Abstract:
The sites of cleavage on the map of the broad-host-range plasmid RK2 (56 kilobases) were determined for the BglII, PstI, and SmaI restriction enzymes, and the determinants for tetracycline and ampicillin resistance were localized. The cleavage sites were clustered at or near the drug resistance genes. To localize regions required for plasmid replication and maintenance in Escherichia coli, we deleted nonessential regions of RK2 by partial digestion with the restriction endonuclease HaeII to produce small derivatives. The smallest stable replicon obtained contained five HaeII fragments of RK2 which total 5.4 kilobases. These fragments were derived from three regions of RK2 that are separated from each other by antibiotic resistance genes. One of these HaeII fragments (0.75 kilobases) has the properties expected of the origin of replication. The outer four fragments, located in two separate regions of RK2, were found to provide, in trans, functions that permit the replication of the HaeII fragment carrying the origin of the replication. These results indicate that at least two plasmid-encoded genes, capable of acting in trans, and a replication origin are required for RK2 replication and maintenance.read more
Citations
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Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.
TL;DR: A broad host range cloning vehicle that can be mobilized at high frequency into Gram-negative bacteria has been constructed from the naturally occurring antibiotic resistance plasmid RK2.
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Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression
Gary S. Ditta,Thomas J. Schmidhauser,Emanuel Yakobson,Peter Lu,Xiao-Wu Liang,Deborah R. Finlay,Donald G. Guiney,Donald R. Helinski +7 more
TL;DR: Derivatives of plasmid pRK290 that are useful for cloning and for analyzing gene expression in a wide variety of Gram-negative bacteria are described.
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Identification and classification of bacterial plasmids.
TL;DR: A bank of hybrid plasmids carrying probes for replicons is constructed and Nick-translational labeling of probe DNA with biotin and colorimetric detection of hybridization is described.
Journal ArticleDOI
The rpoN gene product of Pseudomonas aeruginosa is required for expression of diverse genes, including the flagellin gene.
TL;DR: Findings indicate that the sigma factor encoded by the rpoN gene is used by P. aeruginosa for transcription of a diverse set of genes that specify biosynthetic enzymes, degradative enzymes, and surface components, which include pili and flagella which are required for full virulence of the organism.
Journal ArticleDOI
Regions of broad-host-range plasmid RK2 involved in replication and stable maintenance in nine species of gram-negative bacteria.
TL;DR: The broad-host-range maintenance requirements of plasmid RK2, therefore, are encoded by multiple functions, and the requirement for one or more of these functions varies among gram-negative bacterial species.
References
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Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
Francisco Bolívar,Raymond L. Rodriguez,Patricia J. Greene,Mary C. Betlach,Herbert L. Heyneker,Herbert W. Boyer,Jorge H. Crosa,Stanley Falkow +7 more
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Journal ArticleDOI
Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans.
TL;DR: Results demonstrate that the potentially lethal function specified by fragment B of RK2 is not necessary for replication and that at least one trans-acting function is directly involved in RK 2 replication.
Journal ArticleDOI
R factor transfer in Rhizobium leguminosarum.
TL;DR: R factors of the compatibility class P were transferred between strains of Escherichia coli k12 and Rhizobium leguminosarum and conferred similar levels of antibiotic resistance to those in the corresponding R+ E. coli k 12 hosts, with the exception of carbenicillin resistance which was greatly reduced.
Journal ArticleDOI
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TL;DR: The sequence identifies many of the features responsible for the production of the proteins of the nine known genes of the organism, including initiation and termination sites for the proteins and RNAs.
Journal ArticleDOI
SUPERCOILED CIRCULAR DNA-PROTEIN COMPLEX IN Escherichia coli: PURIFICATION AND INDUCED CONVERSION TO AN OPEN CIRCULAR DNA FORM
TL;DR: Electron microscopic analyses indicate that the decrease in sedimentation rate of the ColE(1)-protein complex after treatment with these various agents is largely owing to an induced transition of ColE (1) DNA from the supercoiled to the open circular state.