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

Technical Report · Tue Jun 01 04:00:00 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
540220* -- Environment
Terrestrial-- Chemicals Monitoring & Transport-- (1990-)
ALKANES
ALKYLATED AROMATICS
AROMATICS
BACTERIA
BIODEGRADATION
BIOTECHNOLOGY
CHEMICAL REACTIONS
CONTAINMENT
DECOMPOSITION
DEHALOGENATION
FEASIBILITY STUDIES
GENETIC ENGINEERING
HERBICIDES
HYDROCARBONS
MICROORGANISMS
OCTANE
ORGANIC COMPOUNDS
PESTICIDES
REMEDIAL ACTION
SOILS
TOLUENE