Containment of genetically engineered organisms after application to subsurface environments. Technical completion report

Technical Report · Tue Jun 01 00:00:00 EDT 1993

OSTI ID:5149077

Crawford, D; Crawford, R; Kellogg, S; Orser, C; Ralston, D

The feasibility of containing genetically engineered bacteria with enhanced dehalogenating properties for in situ bioremediation was investigated. (1) An agarose matrix microbead protocol and a detection system for contained microorganisms or DNA were developed. Multiplex Polymerase Chain Reaction (PCR) allowed tracking of a consortium of encapsulated organisms or several gene targets from a single species. Gene sequences encoding the enzymes responsible for initiating the biodegradation of toluene, octane, and 2,4-D were detected by multiplex PCR and nucleic acid probes from similar to 1-10 biodegradative cells/g soil. Improved DNA extraction methods resulted in PCR reactions detecting similar to 6 cells/g soil. (2) The pcpB gene (for the broad-spectrum detoxicant pentachlorophenol (PCP) hydroxylase) isolated from Flavobacterium sp. strain ATCC 39723 was used in attempts to develop an improved dehalogenating recombinant microorganism for containment experiments.

• Research Organization: Idaho Univ., Moscow, ID (United States). Center for Hazardous Waste Remediation Research
• OSTI ID: 5149077
• Report Number(s): PB-94-144201/XAB; CNN: DI-14-08-0001-G1898
• Country of Publication: United States
• Language: English

Related Subjects

54 ENVIRONMENTAL SCIENCES
BACTERIA
CONTAINMENT
OCTANE
BIODEGRADATION
PESTICIDES
TOLUENE
DEHALOGENATION
FEASIBILITY STUDIES
GENETIC ENGINEERING
HERBICIDES
REMEDIAL ACTION
SOILS
ALKANES
ALKYLATED AROMATICS
AROMATICS
BIOTECHNOLOGY
CHEMICAL REACTIONS
DECOMPOSITION
HYDROCARBONS
MICROORGANISMS
ORGANIC COMPOUNDS
540220* - Environment
Terrestrial- Chemicals Monitoring & Transport- (1990-)