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. 1981 Oct;40(1):197–203. doi: 10.1128/jvi.40.1.197-203.1981

Deletion analysis of the cloned replication origin region from bacteriophage M13.

J M Cleary, D S Ray
PMCID: PMC256609  PMID: 7288922

Abstract

A cloned 270-nucleotide fragment from the origin region of the M13 duplex replicative form DNA confers an M13-dependent replication mechanism upon the plasmid vector pBR322. This M13 insert permits M13 helper-dependent replication of the hybrid plasmid in polA cells which are unable to replicate the pBR322 replicon alone. Using in vitro techniques, we have constructed several plasmids containing deletions in the M13 DNa insert. The endpoints of these deletions have been determined by DNA sequence analysis and correlated with the transformation and replication properties of each plasmid. Characterization of these deletion plasmids allows the following conclusions. (i) The initiation site for M13 viral strand replication is required for helper-dependent propagation of the chimeric plasmid. (ii) A DNA sequence in the M13 insert, localized between 89 and 129 nucleotides from the viral strand initiation site, is necessary for efficient transformation of polA cells. A chimeric plasmid containing the viral strand initiation site, but lacking this additional 40 nucleotide M13 sequence, transforms helper-infected cells at a frequency approximately 10(4)-fold less than that of plasmids containing this additional DNA segment. (iii) The entire M13 complementary strand origin can be deleted without affecting M13-dependent transformation by the hybrid plasmids. We propose a model in which replication of one strand of duplex chimera initiates by nicking at the gene II protein nicking site in the viral strand of the M13 insert, followed by asymmetric single-strand synthesis. Initiation of the complementary strand possibly occurs within plasmid sequences.

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Selected References

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  1. Bahl C. P., Marians K. J., Wu R. A general method for inserting specific DNA sequences into cloning vehicles. Gene. 1976;1(1):81–92. doi: 10.1016/0378-1119(76)90008-1. [DOI] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Cleary J. M., Ray D. S. Replication of the plasmid pBR322 under the control of a cloned replication origin from the single-stranded DNA phage M13. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4638–4642. doi: 10.1073/pnas.77.8.4638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Geider K., Beck E., Schaller H. An RNA transcribed from DNA at the origin of phage fd single strand to replicative form conversion. Proc Natl Acad Sci U S A. 1978 Feb;75(2):645–649. doi: 10.1073/pnas.75.2.645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gray C. P., Sommer R., Polke C., Beck E., Schaller H. Structure of the orgin of DNA replication of bacteriophage fd. Proc Natl Acad Sci U S A. 1978 Jan;75(1):50–53. doi: 10.1073/pnas.75.1.50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gray H. B., Jr, Ostrander D. A., Hodnett J. L., Legerski R. J., Robberson D. L. Extracellular nucleases of Pseudomonas BAL 31. I. Characterization of single strand-specific deoxyriboendonuclease and double-strand deoxyriboexonuclease activities. Nucleic Acids Res. 1975 Sep;2(9):1459–1492. doi: 10.1093/nar/2.9.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Horiuchi K., Zinder N. D. Origin and direction of synthesis of bacteriophage fl DNA. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2341–2345. doi: 10.1073/pnas.73.7.2341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Humphreys G. O., Willshaw G. A., Anderson E. S. A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975 Apr 2;383(4):457–463. doi: 10.1016/0005-2787(75)90318-4. [DOI] [PubMed] [Google Scholar]
  10. Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Klein R. D., Selsing E., Wells R. D. A rapid microscale technique for isolation of recombinant plasmid DNA suitable for restriction enzyme analysis. Plasmid. 1980 Jan;3(1):88–91. doi: 10.1016/s0147-619x(80)90037-2. [DOI] [PubMed] [Google Scholar]
  12. Langridge J., Langridge P., Bergquist P. L. Extraction of nucleic acids from agarose gels. Anal Biochem. 1980 Apr;103(2):264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  13. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  14. Marvin D. A., Hohn B. Filamentous bacterial viruses. Bacteriol Rev. 1969 Jun;33(2):172–209. doi: 10.1128/br.33.2.172-209.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Messing J., Gronenborn B., Müller-Hill B., Hans Hopschneider P. Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3642–3646. doi: 10.1073/pnas.74.9.3642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Meyer T. F., Geider K., Kurz C., Schaller H. Cleavage site of bacteriophage fd gene II-protein in the origin of viral strand replication. Nature. 1979 Mar 22;278(5702):365–367. doi: 10.1038/278365a0. [DOI] [PubMed] [Google Scholar]
  18. Nomura N., Ray D. S. Expression of a DNA strand initiation sequence of ColE1 plasmid in a single-stranded DNA phage. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6566–6570. doi: 10.1073/pnas.77.11.6566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nomura N., Ray D. S. Replication of bacteriophage M13. XV. Location of the specific nick in M13 replicative form II accumulated in Escherichia coli polAex1. J Virol. 1980 Apr;34(1):162–167. doi: 10.1128/jvi.34.1.162-167.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ray D. S., Dueber J. Location of the origin-terminus of the viral strand of the duplex replicative form of bacteriophage G4 DNA. J Mol Biol. 1977 Jul 15;113(4):652–661. doi: 10.1016/0022-2836(77)90228-5. [DOI] [PubMed] [Google Scholar]
  21. Schaller H. The intergenic region and the origins for filamentous phage DNA replication. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):401–408. doi: 10.1101/sqb.1979.043.01.046. [DOI] [PubMed] [Google Scholar]
  22. Schaller H., Uhlmann A., Geider K. A DNA fragment from the origin of single-strand to double-strand DNA replication of bacteriophage fd. Proc Natl Acad Sci U S A. 1976 Jan;73(1):49–53. doi: 10.1073/pnas.73.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Suggs S. V., Ray D. S. Nucleotide sequence of the origin for bacteriophage M13 DNA replication. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):379–388. doi: 10.1101/sqb.1979.043.01.044. [DOI] [PubMed] [Google Scholar]
  24. Suggs S. V., Ray D. S. Replication of bacteriophage M13. XI. Localization of the origin for M13 single-strand synthesis. J Mol Biol. 1977 Feb 15;110(1):147–163. doi: 10.1016/s0022-2836(77)80103-4. [DOI] [PubMed] [Google Scholar]
  25. Zipursky S. L., Reinberg D., Hurwitz J. In vitro DNA replication of recombinant plasmid DNAs containing the origin of progeny replicative form DNA synthesis of phage phi X174. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5182–5186. doi: 10.1073/pnas.77.9.5182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. van Wezenbeek P. M., Hulsebos T. J., Schoenmakers J. G. Nucleotide sequence of the filamentous bacteriophage M13 DNA genome: comparison with phage fd. Gene. 1980 Oct;11(1-2):129–148. doi: 10.1016/0378-1119(80)90093-1. [DOI] [PubMed] [Google Scholar]

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