Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1979 Apr;76(4):1648-52.
doi: 10.1073/pnas.76.4.1648.

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

D H Figurski, D R Helinski

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

D H Figurski et al. Proc Natl Acad Sci U S A. 1979 Apr.

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 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 RK2 replication.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. J Mol Biol. 1963 Mar;6:208-13 - PubMed
    1. Mol Gen Genet. 1977 Apr 29;152(3):175-82 - PubMed
    1. Mol Gen Genet. 1977 Apr 29;152(3):129-35 - PubMed
    1. Biochim Biophys Acta. 1977 Sep 6;478(1):109-13 - PubMed
    1. Science. 1977 Apr 8;196(4286):172-4 - PubMed

Publication types

Substances

LinkOut - more resources