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.
Rumen Bacterium Ruminal Bacterium Prevotella Species Nucleolytic Activity Deoxyribonuclease Activity
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.
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).Google Scholar
Ausubel F.M.:Current Protocols in Molecular Biology. John Wiley & Sons, New York 1987.Google Scholar
Benedik M. J., Strych U.:Serratia marcescens and its extracellular nuclease.FEMS Microbiol. Lett.,165, 1–13 (1998).PubMedCrossRefGoogle Scholar
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).PubMedCrossRefGoogle Scholar
Focareta T., Manning P.A.: Distinguishing between the extracellular DNAases ofVibrio cholerae and development of a transformation system.Mol. Microbiol..5, 2547–2555 (1991).PubMedCrossRefGoogle Scholar
Fomenkov A., Xu S.: Cloning of a gene fromThermus filiformis and characterization of the thermostable nuclease.Gene.163, 109–113 (1995).PubMedCrossRefGoogle Scholar
Hobson P.N.: Rumen bacteria, pp. 133–149 in J.R. Noriss, D.W. Ribbons (Eds):Methods in Microbiology. Academic Press, New York 1969.Google Scholar
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).PubMedGoogle Scholar
Medrano J.F.: DNA extraction from nucleated red blood cells.Biotechniques.8, 43 (1990).PubMedGoogle Scholar
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).PubMedCrossRefGoogle Scholar
Molnarova V., Pristaš P., Javorský P.: Prevalence of CTGCAG recognizing restriction and modification systems in ruminal selenomonads.Anaerobe.5, 37–41 (1999).PubMedCrossRefGoogle Scholar
Rosenthal A.L., Lacks S.A.: Nuclease detection in SDS-polyacrylamide gel electrophoresis.Anal. Biochem.80, 76–90 (1977).PubMedCrossRefGoogle Scholar