Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in <i>trans</i>
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- David H. Figurski
- Department of Biology, University of California at San Diego, La Jolla, California 92093
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- Donald R. Helinski
- Department of Biology, University of California at San Diego, La Jolla, California 92093
Abstract
<jats:p> pRK212.2, a derivative of the broad host range plasmid RK2, contains two <jats:italic>Eco</jats:italic> RI cleavage fragments, A and B, neither of which can replicate by itself in <jats:italic>Escherichia coli</jats:italic> . 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 <jats:italic>E. coli</jats:italic> 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 <jats:italic>E. coli polA</jats:italic> mutants only when a fragment A-bearing helper plasmid is present. These results demonstrate that the potentially lethal function specified by fragment B of RK2 is not necessary for replication and that at least one <jats:italic>trans</jats:italic> -acting function is directly involved in RK2 replication. </jats:p>
Journal
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- Proceedings of the National Academy of Sciences
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Proceedings of the National Academy of Sciences 76 (4), 1648-1652, 1979-04
Proceedings of the National Academy of Sciences
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Keywords
Details 詳細情報について
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- CRID
- 1361981468690885120
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- NII Article ID
- 30016268480
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- ISSN
- 10916490
- 00278424
- http://id.crossref.org/issn/00278424
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- Data Source
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- Crossref
- CiNii Articles