PMID: 377280Apr 1, 1979Paper

Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans

Proceedings of the National Academy of Sciences of the United States of America
D H Figurski, D R Helinski

Abstract

pRK212.2, a derivative of the broad host range plasmid RK2, contains two EcoRI cleavage fragments, A and B, neither of which can replicate by itself in Escherichia coli. Fragment A (41.7 kilobases), but not fragment B (14.4 kilobases), can be cloned by insertion into the unrelated plasmids mini-F and ColE1. Fragment B contains the origin of replication and the ampicillin-resistance determinant of RK2. Transformation of E. coli cells containing the mini-F-fragment A hybrid plasmid with fragment B DNA results in the recircularization and replication of fragment B as a nonmobilizable plasmid (pRK2067) with the copy number and incompatibility properties of RK2. Fragment B cannot be cloned in the absence of fragment A because the latter fragment suppresses a function, specified by fragment B, that results in loss of host cell viability. A small segment (2.4 kilobases) of fragment B that contains the RK2 origin of replication but no longer affects host cell growth in the absence of fragment A had been cloned previously by insertion into a ColE1 plasmid. This hybrid plasmid, designated pRK256, will replicate in E. coli polA mutants only when a fragment A-bearing helper plasmid is present. These results demonstrate that the potentially...Continue Reading

References

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Citations

May 1, 1985·Plant Molecular Biology·P van den ElzenJ Bedbrook
Jan 1, 1987·Plant Molecular Biology·F Rodriguez-QuinonesA Kondorosi
Mar 1, 1985·Plant Molecular Biology·C WaldronS K Malcolm
Aug 1, 1993·Plant Molecular Biology·P A LessardR N Beachy
May 1, 1988·Plant Molecular Biology·S J SandlerP Dunsmuir
Dec 1, 1992·Plant Molecular Biology·T FujiwaraR N Beachy
Nov 1, 1991·Plant Molecular Biology·R HightowerP Dunsmuir
Nov 1, 1990·Molecular & General Genetics : MGG·G BlancoM Tortolero
Oct 17, 1994·Molecular & General Genetics : MGG·V VenturiP J Weisbeek
Nov 1, 1995·Molecular & General Genetics : MGG·I D'hoogheJ Vanderleyden
Jan 1, 1983·Molecular & General Genetics : MGG·K Tanimoto, T Iino
Sep 1, 1987·Molecular & General Genetics : MGG·A Contreras, J Casadesús
Jan 1, 1983·Molecular & General Genetics : MGG·C A Smith, C M Thomas
Jan 1, 1984·Molecular & General Genetics : MGG·P T BarthD H Figurski
Oct 1, 1987·Molecular & General Genetics : MGG·H M Fischer, H Hennecke
Aug 1, 1989·Molecular & General Genetics : MGG·T L AldrichA M Chakrabarty
Mar 1, 1986·Molecular & General Genetics : MGG·K Poole, R E Hancock

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