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. 1981 Jan 10;9(1):85–93. doi: 10.1093/nar/9.1.85

Temperature dependence of the joining by T4 DNA ligase of termini produced by type II restriction endonucleases.

L Ferretti, V Sgaramella
PMCID: PMC326670  PMID: 6259621

Abstract

The temperature dependence of the T4 DNA ligase-catalyzed joining of plasmid DNA linearized by the action of HaeIII, EcoRI and PstI restriction endonucleases has been investigated by electron microscopy analysis. The extent of joining is maximal at 4 degrees and decreases with increasing temperatures following sigmoid-like curves. The temperature at which 50% of the maximal reaction is still observable increases going from DNA termini without single-stranded overlaps (produced by HaeIII) to termini with four nucleotides overlap, composed only by two A and two T (produced by EcoRI) to termini with four nucleotide overlap, composed by A, T, G and C (produced by PstI).

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Selected References

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  1. Bittner M., Burke R. L., Alberts B. M. Purification of the T4 gene 32 protein free from detectable deoxyribonuclease activities. J Biol Chem. 1979 Oct 10;254(19):9565–9572. [PubMed] [Google Scholar]
  2. 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]
  3. Dugaiczyk A., Boyer H. W., Goodman H. M. Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures. J Mol Biol. 1975 Jul 25;96(1):171–184. doi: 10.1016/0022-2836(75)90189-8. [DOI] [PubMed] [Google Scholar]
  4. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Higgins N. P., Cozzarelli N. R. DNA-joining enzymes: a review. Methods Enzymol. 1979;68:50–71. doi: 10.1016/0076-6879(79)68006-0. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Mertz J. E., Davis R. W. Cleavage of DNA by R 1 restriction endonuclease generates cohesive ends. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3370–3374. doi: 10.1073/pnas.69.11.3370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Miller L. K., Fried M. Construction of infectious polyoma hybrid genomes in vitro. Nature. 1976 Feb 19;259(5544):598–601. doi: 10.1038/259598a0. [DOI] [PubMed] [Google Scholar]
  10. Mottes M., Morandi C., Cremaschi S., Sgaramella V. Restoration by T4 ligase of DNA sequences sensitive to "flush" cleaving restriction enzyme. Nucleic Acids Res. 1977 Jul;4(7):2467–2475. doi: 10.1093/nar/4.7.2467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Murray N. E., Bruce S. A., Murray K. Molecular cloning of the DNA ligase gene from bacteriophage T4. II. Amplification and preparation of the gene product. J Mol Biol. 1979 Aug 15;132(3):493–505. doi: 10.1016/0022-2836(79)90271-7. [DOI] [PubMed] [Google Scholar]
  12. Roberts R. J. Restriction and modification enzymes and their recognition sequences. Gene. 1978 Nov;4(3):183–194. doi: 10.1016/0378-1119(78)90017-3. [DOI] [PubMed] [Google Scholar]
  13. Sgaramella V., Ehrlich S. D. Use of the T4 polynucleotide ligase in the joining of flush-ended DNA segments generated by restriction endonucleases. Eur J Biochem. 1978 May 16;86(2):531–537. doi: 10.1111/j.1432-1033.1978.tb12336.x. [DOI] [PubMed] [Google Scholar]
  14. Tait R. C., Rodriguez R. L., West R. W., Jr The rapid purification of T4 DNA ligase from a lambda T4 lig lysogen. J Biol Chem. 1980 Feb 10;255(3):813–815. [PubMed] [Google Scholar]
  15. Vosberg H. P. Molecular cloning of DNA. An introduction into techniques and problems. Hum Genet. 1977 Dec 29;40(1):1–72. doi: 10.1007/BF00280831. [DOI] [PubMed] [Google Scholar]

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