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. 1990 May;172(5):2762–2764. doi: 10.1128/jb.172.5.2762-2764.1990

Second-site revertants of the P1 copN22 copy mutant.

B J Froehlich 1, J R Scott 1
PMCID: PMC208922  PMID: 2185232

Abstract

Miniplasmids with the P1 copN22 mutation have a copy number about seven times that of the wild type. Selection for reduced copy number from this plasmid led to the isolation of second-site pseudorevertants, called poc mutants. DNA sequence analysis showed that all six independent poc mutants have a single base change in the same codon of the repA gene. This implicates the amino acid at this location, either directly or indirectly, in interactions important for copy number control.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abeles A. L., Friedman S. A., Austin S. J. Partition of unit-copy miniplasmids to daughter cells. III. The DNA sequence and functional organization of the P1 partition region. J Mol Biol. 1985 Sep 20;185(2):261–272. doi: 10.1016/0022-2836(85)90402-4. [DOI] [PubMed] [Google Scholar]
  2. Abeles A. L. P1 plasmid replication. Purification and DNA-binding activity of the replication protein RepA. J Biol Chem. 1986 Mar 15;261(8):3548–3555. [PubMed] [Google Scholar]
  3. Abeles A. L., Snyder K. M., Chattoraj D. K. P1 plasmid replication: replicon structure. J Mol Biol. 1984 Mar 5;173(3):307–324. doi: 10.1016/0022-2836(84)90123-2. [DOI] [PubMed] [Google Scholar]
  4. Austin S. J., Mural R. J., Chattoraj D. K., Abeles A. L. Trans- and cis-acting elements for the replication of P1 miniplasmids. J Mol Biol. 1985 May 25;183(2):195–202. doi: 10.1016/0022-2836(85)90212-8. [DOI] [PubMed] [Google Scholar]
  5. Baumstark B. R., Lowery K., Scott J. R. Location by DNA sequence analysis of cop mutations affecting the number of plasmid copies of prophage P1. Mol Gen Genet. 1984;194(3):513–516. doi: 10.1007/BF00425567. [DOI] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chattoraj D. K., Mason R. J., Wickner S. H. Mini-P1 plasmid replication: the autoregulation-sequestration paradox. Cell. 1988 Feb 26;52(4):551–557. doi: 10.1016/0092-8674(88)90468-0. [DOI] [PubMed] [Google Scholar]
  8. Chattoraj D. K., Snyder K. M., Abeles A. L. P1 plasmid replication: multiple functions of RepA protein at the origin. Proc Natl Acad Sci U S A. 1985 May;82(9):2588–2592. doi: 10.1073/pnas.82.9.2588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chattoraj D., Cordes K., Abeles A. Plasmid P1 replication: negative control by repeated DNA sequences. Proc Natl Acad Sci U S A. 1984 Oct;81(20):6456–6460. doi: 10.1073/pnas.81.20.6456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Froehlich B. J., Scott J. R. A single amino acid difference between Rep proteins of P1 and P7 affects plasmid copy number. Plasmid. 1988 Mar;19(2):121–133. doi: 10.1016/0147-619x(88)90051-0. [DOI] [PubMed] [Google Scholar]
  13. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  14. Kahn M. L., Figurski D., Ito L., Helinski D. R. Essential regions for replication of a stringent and a relaxed plasmid in Escherichia coli. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):99–103. doi: 10.1101/sqb.1979.043.01.015. [DOI] [PubMed] [Google Scholar]
  15. Pal S. K., Mason R. J., Chattoraj D. K. P1 plasmid replication. Role of initiator titration in copy number control. J Mol Biol. 1986 Nov 20;192(2):275–285. doi: 10.1016/0022-2836(86)90364-5. [DOI] [PubMed] [Google Scholar]
  16. Prentki P., Chandler M., Caro L. Replication of prophage P1 during the cell cycle of Escherichia coli. Mol Gen Genet. 1977 Mar 28;152(1):71–76. doi: 10.1007/BF00264942. [DOI] [PubMed] [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Scott J. R. A turbid plaque-forming mutant of phage P1 that cannot lysogenize Escherichia coli. Virology. 1974 Dec;62(2):344–349. doi: 10.1016/0042-6822(74)90397-3. [DOI] [PubMed] [Google Scholar]
  19. Scott J. R., Kropf M. M., Padolsky L., Goodspeed J. K., Davis R., Vapnek D. Mutants of plasmid prophage P1 and elevated copy number: isolation and characterization. J Bacteriol. 1982 Jun;150(3):1329–1339. doi: 10.1128/jb.150.3.1329-1339.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Uhlin B. E., Nordström K. R plasmid gene dosage effects in Escherichia coli K-12: copy mutants of the R plasmic R1drd-19. Plasmid. 1977 Nov;1(1):1–7. doi: 10.1016/0147-619x(77)90003-8. [DOI] [PubMed] [Google Scholar]

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