Molecular biology of environmental aromatic hydrocarbons. Progress report, September 1, 1985-July 31, 1986

Technical Report · Fri Aug 01 04:00:00 EDT 1986

OSTI ID:5278769

Weiss, S B

Recent studies in our laboratory have indicated that short DNA oligonucleotides can be used as targets for alkylation by activated benzo(a)pyrene diol epoxide (BPDE) to examine the effect of PAH-induced mutagenesis. The alkylated oligomers are ligated into viral DNA and this construct is used to tranfect Escherichia coli. Progeny virus are selected from agar plates, amplified, and the viral DNA isolated is sequenced at the site of oligomer insertion. A scheme for the assembly of synthetic DNA oligonucleotides has been devised such that: (a) the site of alkylation in the oligomer is specific for a single deoxynucleotide species; and (b) the position of the adduct(s) in the oligomer can be varied. At the present stage of our work, we have prepared four synthetic duplex DNA oligomers. These oligomers are 32 base pairs in length and contain either a single or double dG residue in each strand separated by 4 or 12 base pairs (see text in this report). When these oligomers are reacted with (+)BPDE, only the dG residues form adducts. Transfection of E. coli with the four different assembled oligomers alkylated with (+)BPDE gives rise to the production of progency viral DNAs which primarily show the restoration of the original ologomer DNA sequence, but also, the appearance of a large deletion at the site of alkylation. The number of progeny viral DNAs sequenced to date are too few to determine a reliable mutation frequency. Construction of DNA oligomers containing other alkylated residues, e.g., dA and dC, is in progress. Preliminary attempts to detect BPDE-induced enzymes that participate in the repair of PAH-damaged DNA are also described. 27 refs., 1 fig.

• Research Organization: Chicago Univ., IL (USA)
• DOE Contract Number: FG02-86ER60394
• OSTI ID: 5278769
• Report Number(s): DOE/ER/60394-1; ON: DE86015797
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
AROMATICS
BENZOPYRENE
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CHEMISTRY
CONDENSED AROMATICS
DAYS LIVING RADIOISOTOPES
DNA ADDUCTS
DNA REPAIR
DNA SEQUENCING
DNA-ASE
ENDONUCLEASES
ENZYMES
EPOXIDES
ESTERASES
EVEN-ODD NUCLEI
FRACTIONATION
HYDROCARBONS
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
NUCLEI
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PHOSPHODIESTERASES
RADIOISOTOPES
RECOVERY
REPAIR
SEPARATION PROCESSES
STRUCTURAL CHEMICAL ANALYSIS
SULFUR 35
SULFUR ISOTOPES
TRACER TECHNIQUES