Abstract
Extracellular non-specific nucleases were observed in some strains belonging to the ruminal species of the genusPrevotella, mostlyP. brevis andP. bryantii. The nuclease fromP. bryantii appeared to be extracellular; it mediates the degradation of the supercoiled plasmid DNAvia an open circle intermediate. The cleavage is not site specific although a preference for certain cleavage sites does seem to exist. Our attempts to clone the wild-typeP. bryantii B14 nuclease inE. coli strain ER1992 that reports on the DNA damage sustained, were unsuccessful probably due to excessive intracellular nuclease activity that killed the cells bearing the gene for the nuclease. On the other hand, the nuclease from a related strain TC1-1, which has a less active enzyme of the same type, was successfully cloned.
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References
Accetto T., Avguštin G.: Deoxyribonuclease activities of rumen bacteria from the genusPrevotella.Res. Rep. Biotech. Fac. Univ. Ljubljana (Agruc. Issue, Zootech.) 74, 83–88 (1999).
Ausubel F.M.:Current Protocols in Molecular Biology. John Wiley & Sons, New York 1987.
Benedik M. J., Strych U.:Serratia marcescens and its extracellular nuclease.FEMS Microbiol. Lett.,165, 1–13 (1998).
Blank A., Sugiyama R.H., Dekker C.A.: Activity staining of nucleolytic enzymes after SDS-polyacrylamide gel electrophoresis: use of aqueous isopropanol to remove detergent from gels.Anal. Biochem.,120, 267–275 (1982).
Flint H.J., Thomson A.M.: Deoxyribonuclease activity in rumen bacteria.Lett. Appl. Microbiol.,11, 18–21 (1990).
Focareta T., Manning P.A.: Distinguishing between the extracellular DNAases ofVibrio cholerae and development of a transformation system.Mol. Microbiol..5, 2547–2555 (1991).
Fomenkov A., Xu S.: Cloning of a gene fromThermus filiformis and characterization of the thermostable nuclease.Gene.163, 109–113 (1995).
Hobson P.N.: Rumen bacteria, pp. 133–149 in J.R. Noriss, D.W. Ribbons (Eds):Methods in Microbiology. Academic Press, New York 1969.
Lee F.S., Forsberg C.W., Gibbins A.M.: Type II restriction-modification system and endonuclease from ruminal bacteriumFibrobacter succinogenes S85.J. Bacteriol..174, 5275–5283 (1992).
Medrano J.F.: DNA extraction from nucleated red blood cells.Biotechniques.8, 43 (1990).
Muro-Pastor A.M., Flores E., Herrero A.: Identification, genetic analysis and characterization of sugar-non-specific nuclease from the cyanobacteriumAnabaena sp. PCC 7120.Mol. Microbiol..6, 3021–3030 (1992).
Molnarova V., Pristaš P., Javorský P.: Prevalence of CTGCAG recognizing restriction and modification systems in ruminal selenomonads.Anaerobe.5, 37–41 (1999).
Rosenthal A.L., Lacks S.A.: Nuclease detection in SDS-polyacrylamide gel electrophoresis.Anal. Biochem. 80, 76–90 (1977).
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Accetto, T., Avguštin, G. Non-specific DNAases from the rumen bacteriumPrevotella bryantii . Folia Microbiol 46, 31–35 (2001). https://doi.org/10.1007/BF02825880
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DOI: https://doi.org/10.1007/BF02825880