Distribution and molecular characterization of dissolved DNA in aquatic environments
The distribution of dissolved DNA in oceanic, estuarine and freshwater environments in southwest Florida and the Gulf of Mexico was determined by using a method for the measurement of dissolved DNA based on the fluorescence of Hoechst 33258-DNA complexes. A wide range of molecular weights (determined by agarose gel electrophoresis) was found for extracellular DNA concentrated from various aquatic environments. A model system for probing extracellular DNA from aquatic environments was developed using the plasmid pS{beta}TK2.0 containing the herpes simplex thymidine kinase (TK) gene. Plasmid DNA and the TK gene fragment added to artificial seawater were concentrated and probed with ({sup 35}S) labelled TK to establish percent recovery and detection limits for the method. The degradation of plasmid DNA added to a natural seawater sample was monitored over a 36 h period by probing with the TK gene probe. Intact plasmid was detected for up to 4 h and DNA hybridizable to the TK probe was detected for up to 24 h. These methods were used to probe for the TK gene in environmental samples of extracellular DNA. Hybridization to the TK probe was detected in both freshwater and estuarine samples.
- Research Organization:
- University of South Florida, Tampa, FL (USA)
- OSTI ID:
- 5283592
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA
BIOASSAY
PHOSPHOTRANSFERASES
GENES
DNA HYBRIDIZATION
ELECTROPHORESIS
ESTUARIES
FLORIDA
FRESH WATER
GULF OF MEXICO
HYBRIDIZATION
MATHEMATICAL MODELS
MEASURING METHODS
MOLECULAR WEIGHT
PLASMIDS
SEAWATER
SPATIAL DISTRIBUTION
SULFUR 35
TRACER TECHNIQUES
ATLANTIC OCEAN
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CARIBBEAN SEA
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
DISTRIBUTION
ENZYMES
EVEN-ODD NUCLEI
FEDERAL REGION IV
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
NORTH AMERICA
NUCLEI
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
RADIOISOTOPES
SEAS
SULFUR ISOTOPES
SURFACE WATERS
TRANSFERASES
USA
WATER
540310* - Environment
Aquatic- Basic Studies- (1990-)