Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1976 Apr;73(4):1068–1072. doi: 10.1073/pnas.73.4.1068

Construction and propagation of a defective simian virus 40 genome bearing an operator from bacteriophage lambda.

A L Nussbaum, D Davoli, D Ganem, G C Fareed
PMCID: PMC430201  PMID: 177971

Abstract

A 2400 base pair DNA segment containing the leftward operator (OL) of phage lambda was covalently joined in vitro to a fragment of simian virus 40 (SV40) DNA harboring the SV40 replication origin. The recombinant molecule was propagated in the presence of helper wild-type SV40 DNA in monkey kidney cells and partially cloned by an infectious center procedure. After propagation in monkey cells and purification, the hybrid DNA could be distinguished from wild-type SV40 DNA by its shortened length (about 80% that of SV40), specific hybridization to denatured lambda DNA immobilized on filters, specific affinity for lambda repressor, and preservation of a large part (about 2300 base pairs) of the lambda immunity region as determined by restriction nuclease cleavage patterns and electron microscopic heteroduplex analysis. These results indicate that defective SV40 replicons can serve as vectors for propagating foreign DNA in mammalian cells.

Full text

PDF
1068

Images in this article

1069Image
on p.1069
1070Image
on p.1070
1070Image
on p.1070
1070Image
on p.1070
1071Image
on p.1071
1071Image
on p.1071
1071Image
on p.1071
1072Image
on p.1072

Selected References

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

  1. Abrahams P. J., Mulder C., Van De Voorde A., Warnaar S. O., van der Eb A. J. Transformation of primary rat kidney cells by fragments of simian virus 40 DNA. J Virol. 1975 Oct;16(4):818–823. doi: 10.1128/jvi.16.4.818-823.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allet B. Fragments produced by cleavage of lambda deoxyribonucleic acid with the Hemophilus parainfluenzae restriction enzyme Hpa II. Biochemistry. 1973 Sep 25;12(20):3972–3977. doi: 10.1021/bi00744a029. [DOI] [PubMed] [Google Scholar]
  3. Allet B., Jeppesen P. G., Katagiri K. J., Delius H. Mapping the DNA fragments produced by cleavage by lambda DNA with endonuclease RI. Nature. 1973 Jan 12;241(5385):120–123. doi: 10.1038/241120a0. [DOI] [PubMed] [Google Scholar]
  4. Allet B., Solem R. Separation and analysis of promoter sites in bacteriophage lambda DNA by specific endonucleases. J Mol Biol. 1974 Jan 5;85(4):475–484. doi: 10.1016/0022-2836(74)90310-6. [DOI] [PubMed] [Google Scholar]
  5. Cameron J. R., Panasenko S. M., Lehman I. R., Davis R. W. In vitro construction of bacteriophage lambda carrying segments of the Escherichia coli chromosome: selection of hybrids containing the gene for DNA ligase. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3416–3420. doi: 10.1073/pnas.72.9.3416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Davoli D., Fareed G. C. Formation of reiterated simian virus 40 DNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):137–146. doi: 10.1101/sqb.1974.039.01.019. [DOI] [PubMed] [Google Scholar]
  7. Fareed G. C., Byrne J. C., Martin M. A. Triplication of a unique genetic segment in a simian virus 40-like virus of human origin and evolution of new viral genomes. J Mol Biol. 1974 Aug 5;87(2):275–288. doi: 10.1016/0022-2836(74)90149-1. [DOI] [PubMed] [Google Scholar]
  8. Fried M., Griffin B. E., Lund E., Robberson D. L. Polyoma virus--a study of wild-type, mutant and defective DNAs. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):45–52. doi: 10.1101/sqb.1974.039.01.009. [DOI] [PubMed] [Google Scholar]
  9. Fried M. Isolation and partial characterization of different defective DNA molecules derived from polyoma virus. J Virol. 1974 May;13(5):939–946. doi: 10.1128/jvi.13.5.939-946.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ganem D., Nussbaum A. L., Davoli D., Fareed G. C. Isolation, propagation and characterization of replication requirements of reiteration mutants of Simian Virus 40. J Mol Biol. 1976 Feb 15;101(1):57–83. doi: 10.1016/0022-2836(76)90066-8. [DOI] [PubMed] [Google Scholar]
  11. Ganem D., Nussbaum A. L., Davoli D., Fareed G. C. Propagation of a segment of bacteriophage lamda-DNA in monkey cells after covalent linkage to a defective simian virus 40 genome. Cell. 1976 Mar;7(3):349–359. doi: 10.1016/0092-8674(76)90164-1. [DOI] [PubMed] [Google Scholar]
  12. Greenfield L., Simpson L., Kaplan D. Conversion of closed circular DNA molecules to single-nicked molecules by digestion with DNAase I in the presence of ethidium bromide. Biochim Biophys Acta. 1975 Oct 15;407(3):365–375. doi: 10.1016/0005-2787(75)90104-5. [DOI] [PubMed] [Google Scholar]
  13. Khoury G., Fareed G. C., Berry K., Martin M. A., Lee T. N., Nathans D. Characterization of a rearrangement in viral DNA: mapping of the circular simian virus 40-like DNA containing a triplication of a specific one-third of the viral genome. J Mol Biol. 1974 Aug 5;87(2):289–301. doi: 10.1016/0022-2836(74)90150-8. [DOI] [PubMed] [Google Scholar]
  14. Lai C. J., Nathans D. Deletion mutants of simian virus 40 generated by enzymatic excision of DNA segments from the viral genome. J Mol Biol. 1974 Oct 15;89(1):179–193. doi: 10.1016/0022-2836(74)90169-7. [DOI] [PubMed] [Google Scholar]
  15. Lai C. J., Nathans D. Non-specific termination of simian virus 40 DNA replication. J Mol Biol. 1975 Sep 5;97(1):113–118. doi: 10.1016/s0022-2836(75)80026-x. [DOI] [PubMed] [Google Scholar]
  16. Lee T. N., Brockman W. W., Nathans D. Evolutionary variants of simian virus 40: cloned substituted variants containing multiple initiation sites for DNA replication. Virology. 1975 Jul;66(1):53–69. doi: 10.1016/0042-6822(75)90178-6. [DOI] [PubMed] [Google Scholar]
  17. Maniatis T., Ptashne M. Structure of the lambda operators. Nature. 1973 Nov 16;246(5429):133–136. doi: 10.1038/246133a0. [DOI] [PubMed] [Google Scholar]
  18. Morrow J. F., Cohen S. N., Chang A. C., Boyer H. W., Goodman H. M., Helling R. B. Replication and transcription of eukaryotic DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1974 May;71(5):1743–1747. doi: 10.1073/pnas.71.5.1743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Murray N. E., Murray K. Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature. 1974 Oct 11;251(5475):476–481. doi: 10.1038/251476a0. [DOI] [PubMed] [Google Scholar]
  20. Old R., Murray K., Boizes G. Recognition sequence of restriction endonuclease III from Hemophilus influenzae. J Mol Biol. 1975 Feb 25;92(2):331–339. doi: 10.1016/0022-2836(75)90232-6. [DOI] [PubMed] [Google Scholar]
  21. Perricaudet M., Tiollais P. Defective bacteriophage lambda chromosome, potential vector for DNA fragments obtained after cleavage by Bacillus amyloliquefaciens endonuclease (BamI). FEBS Lett. 1975 Aug 1;56(1):7–11. doi: 10.1016/0014-5793(75)80099-8. [DOI] [PubMed] [Google Scholar]
  22. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  23. Wilson G. A., Young F. E. Isolation of a sequence-specific endonuclease (BamI) from Bacillus amyloliquefaciens H. J Mol Biol. 1975 Sep 5;97(1):123–125. doi: 10.1016/s0022-2836(75)80028-3. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES