SEDL / STP / STP1115-EB / STP19524S

³²P-Postlabeling for DNA Adduct Determination in Plants

Lower, WR
Group leader, Environmental Trace Substances Research Center, University of Missouri, Columbia, MO

Ireland, FA
Research specialist, Dixon Springs Agricultural Center, University of Illinois, Simpson, IL

Judy, BM
Research specialist, University of Missouri, Columbia, MO

Pages: 12    Published: Jan 1991

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Abstract

Barley (Hordeum vulgare L.) and Australian saltbush (Atriplex semibaccata)plants and deoxyribonucleic acid (DNA) isolated from barley were exposed to the known carcinogen/mutagen, N-methyl-N-nitrosourea (MNU). DNA was isolated, digested to a mixture of deoxyribonucleoside 5'-monophosphates, postlabeled with [γ-³²P] ATP, and separated on polyethyleneimine (PEI)-cellulose thin layer chromatography plates. Autoradiograms of the chromatograms showed the presence of normal DNA constituents as well as additional spots not present on autoradiograms of untreated DNA. These data suggest that DNA adducts are formed in growing plants and in plant DNA exposed to a carcinogen. In addition, a discussion of the ³²P-postlabeling procedure and similarities and differences encountered using plants and plant DNA are addressed.