Abstract
A method is described which allows a large number of bacterial strains to be rapidly and easily screened for the presence of site-specific endonucleases. The method involves selective permeabilization of the bacterial cell and analysis of the exuded material. Type II restriction endonucleases from cyanobacteria and Gram-negative eubacteria have been detected and new enzymes have been found. The method should be widely applicable and easy to modify for use in genera other than those tested. Three-site-specific endonuclease activities, detected by this method in Aphanothece halophytica PCC 7412, were purified and their recognition and cleavage specificities were determined AhaI and AhaII recognise and cleave the same DNA sequences as CauII and AcyI respectively; the specificity of AhaIII (TTT↓AAA) has been reported previously (Whitehead and Brown, 1982, FEBS Letters 143:296–300).
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Abbreviations
- Brij-58:
-
20 cetyl ether
- Pu:
-
purine nucleoside
- Py:
-
pyrimidine nucleoside
References
Arber W (1979) Promotion and limitation of genetic exchange. Science 205:361–365
Brown NL, Smith M (1980) A general method for defining restriction enzyme cleavage and recognition sites. In: Moldave K, Grossman L (eds) Methods in enzymology, vol 65. Academic Press, New York London, pp 391–404
Brown NL, McClelland M, Whitehead PR (1980) HgiAI: a restriction endonuclease from Herpetosiphon giganteus HP1023. Gene 9:46–68
Duncan CH, Wilson GA, Young FE (1978) Biochemical and genetic properties of site-specific restriction endonucleases in Bacillus globigii. J Bacteriol 134:338–344
Fuchs C, Rosenwald EC, Honigman A, Szybalski W (1980) Identification of palindromic sequences recognised by restriction endonucleases, as based on the tabularized sequencing data for seven viral and plasmid DNAs. Gene 10:357–370
Godson GN, Sinsheimer RL (1967) Lysis of Escherichia coli with a neutral detergent. Biochem Biophys Acta 149:476–488
Greene PJ, Heyneker HL, Bolivar F, Rodriguez RL, Betlach MC, Covarrubias AA, Backman K, Russell DJ, Tait R, Boyer HW (1978) A general method for purifying restriction enzymes. Nucleic Acids Res 5:2373–2380
Holt JG, Lewin RA (1968) Herpetosiphon auranticus gen, et sp. n., a new filamentous gliding organism. J Bacteriol 95:2407–2408
Hopwood DA (1981) Genetic studies with bacterial protoplasts. Ann Rev Microbiol 35:237–272
Mayer H, Reichenbach H (1978) Restriction endonucleases: general survey procedure and survey of gliding bacteria. J Bacteriol 136:708–713
Misra TK, Brown NL, Fritzinger DC, Pridmore RD, Barnes WM, Haberstroh L, Silver S (1984) Mercuric ion-resistance operons of plasmid R100 and transposon Tn501: the beginning of the operon including the regulatory region and the first two structural genes. Proc Natl Acad Sci USA 81:5975–5979
Modrich P, Roberts RJ (1982) Type-II restriction modification enzymes. In: Linn S, Roberts RJ (eds) Nucleases. Cold Spring Harbor Laboratory, New York, pp 109–154
Purvis IJ, Moscley BEB (1983) Isolation and characterisation of DraI, a type II restriction endonuclease recognising a sequence containing only A:T basepairs, and inhibition of its activity by UV irradiation of substrate DNA. Nucleic Acids Res 11:5467–5474
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61
Roberts RJ (1984) Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res 12:r167-r204
Sanger F, Coulson AR, Barrell BG, Smith AJH, Coe BA (1980) Cloning in single stranded DNA as an aid to rapid DNA sequencing. J Mol Biol 143:161–178
Sharp PA, Sugden B, Sambrook J (1973) Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose — ethidium bromide electrophoresis. Biochemistry 12:3055–3063
Smith DI, Blattner FR, Davies J (1976) The isolation and partial characterisation of a new restriction endonuclease from Providencia stuartii. Nucleic Acids Res 2:343–353
Smith HO, Wilcox KW (1970) A restriction enzyme from Haemophilus influenzae I Purification and general properties. J Mol Biol 51:379–391
Staden R (1977) Sequence data handling by computer. Nucleic Acids Res 4:4037–4051
Walsby AE, Brooker MJ (1980) Changes in buoyancy of a plantonic blue-green alga in response to light intensity. Brit Phycol J 15:311–319
Whitehead PR, Brown NL (1982) AhaIII: a restriction endonuclease with a recognition sequence containing only A:T base pairs. FEBS letters 143:296–300
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Whitehead, P.R., Brown, N.L. A simple and rapid method for screening bacteria for type II restriction endonucleases: enzymes in Aphanothece halophytica . Arch. Microbiol. 141, 70–74 (1985). https://doi.org/10.1007/BF00446742
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DOI: https://doi.org/10.1007/BF00446742