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Nucleic Acids Res. 1987 October 12; 15(19): 7677–7687.
PMCID: PMC306300

Alteration of the specificity of PvuII restriction endonuclease.


The restriction endonuclease PvuII which cleaves the sequence CAGCTG, at the position indicated by the arrow, was found to decrease its substrate specificity in the presence of organic solvents. Thirty-three sites, that we have named PvuII sites, were identified on the nucleotide sequence of pBR322 DNA. The new recognition sequences cleaved in pBR322 DNA, at the positions indicated by the arrows, were shown to be AAGCTG, GAGCTG, CNGCTG, CANCTG, CAGNTG, CAGCNG, CAGCTC and CAGCTT. (TAGCTG and the complementary sequence CAGCTA are not present in pBR322 DNA). From these recognition sequences, we deduced that PvuII activity recognizes and cleaves degenerate sequences which differ from the standard PvuII sequence CAGCTG at only one of the recognition site. Any substitution can occur at any one of the six positions in the hexanucleotide sequence. The optimum incubation medium for PvuII activity was found to be: 10-50 mM Tris-HCl, pH 8.5, 12-15 mM MgCl2, 50 mM NaCl, 10% ethanol + 10% dimethylsulfoxide (DMSO).

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Selected References

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  • Nathans D, Smith HO. Restriction endonucleases in the analysis and restructuring of dna molecules. Annu Rev Biochem. 1975;44:273–293. [PubMed]
  • Roberts RJ. Restriction endonucleases. CRC Crit Rev Biochem. 1976 Nov;4(2):123–164. [PubMed]
  • Polisky B, Greene P, Garfin DE, McCarthy BJ, Goodman HM, Boyer HW. Specificity of substrate recognition by the EcoRI restriction endonuclease. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3310–3314. [PubMed]
  • Heininger K, Hörz W, Zachau HG. Specificity of cleavage by restriction nuclease from Bacillus subtilis. Gene. 1977 Jul;1(5-6):291–303. [PubMed]
  • Hsu M, Berg P. Altering the specificity of restriction endonuclease: effect of replacing Mg2+ with Mn2+. Biochemistry. 1978 Jan 10;17(1):131–138. [PubMed]
  • Tikchonenko TI, Karamov EV, Zavizion BA, Naroditsky BS. EcoRI activity: enzyme modification or activation of accompanying endonuclease? Gene. 1978 Nov;4(3):195–212. [PubMed]
  • Clarke CM, Hartley BS. Purification, properties and specificity of the restriction endonuclease from Bacillus stearothermophilus. Biochem J. 1979 Jan 1;177(1):49–62. [PubMed]
  • Malyguine E, Vannier P, Yot P. Alteration of the specificity of restriction endonucleases in the presence of organic solvents. Gene. 1980 Jan;8(2):163–177. [PubMed]
  • Nasri M, Thomas D. Relaxation of recognition sequence of specific endonuclease HindIII. Nucleic Acids Res. 1986 Jan 24;14(2):811–821. [PMC free article] [PubMed]
  • Halford SE, Lovelady BM, McCallum SA. Relaxed specificity of the EcoRV restriction endonuclease. Gene. 1986;41(2-3):173–181. [PubMed]
  • Kolesnikov VA, Zinoviev VV, Yashina LN, Karginov VA, Baclanov MM, Malygin EG. Relaxed specificity of endonuclease BamH1 as determined by identification or recognition sites in SV 40 and pBR 322 DNAs. FEBS Lett. 1981 Sep 14;132(1):101–104. [PubMed]
  • George J, Chirikjian JG. Sequence-specific endonuclease BamHI: relaxation of sequence recognition. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2432–2436. [PubMed]
  • Nasri M, Sayadi S, Thomas D. Relaxation of PvuII recognition sequence. FEBS Lett. 1985 Jun 3;185(1):101–104. [PubMed]
  • Sutcliffe JG. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. [PubMed]
  • Maxam AM, Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. [PubMed]
  • Gingeras TR, Greenough L, Schildkraut I, Roberts RJ. Two new restriction endonucleases from Proteus vulgaris. Nucleic Acids Res. 1981 Sep 25;9(18):4525–4536. [PMC free article] [PubMed]
  • Roberts RJ, Myers PA, Morrison A, Murray K. A specific endonuclease from Arthrobacter luteus. J Mol Biol. 1976 Mar 25;102(1):157–165. [PubMed]

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