Analysis of gene expression following low dose {gamma}-irradiation

Journal Article · Sat Dec 31 04:00:00 EST 1994 · Environmental and Molecular Mutagenesis

OSTI ID:91985

Thurston, S J; Groch, K; Brooks, A; Saffer, J ^[1]

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1. Pacific Northwest Laboratory, Richland, WA (United States)

Low levels of exposure to physical and chemical carcinogens induce repair enzymes which may alter the shape of the dose-response relationship of subsequent exposures. The mechanisms involved in this adaptive response need to be understood to define its potential influence on health risk. To perform a comprehensive analysis of changes in gene expression due to radiation, we have begun to utilize the differential display PCR method. A subset of mRNAs are transcribed to cDNA using a primer that anneals to the poly(A) tail plus two additional bases (e.g., 5{prime}-T{sub 12}CC would define those RNAs that end with GGAn). These cDNAs are then amplified by PCR using a short arbitrary upstream primer resulting in a distinctly sized fragment from each cDNA. In comparing exposed and control cells, a change in the amount of any resulting band would indicate that the mRNA represented by the band has altered expression. Exponentially growing CHO-K1 cell were exposed to 0, 1, 10, and 100 cGy {sup 60}Co {gamma}-irradiation. RNA isolated from these cells has been screened for differential expression. Approximately 1/4 of the mRNAs within the cells have been examined without revealing candidate genes with altered expression. We have shown that responsiveness to other insults can be identified by ddPCR and that candidate can rapidly be cloned, sequenced and confirmed by Northern analysis.

• OSTI ID: 91985
• Report Number(s): CONF-9405324--
• Journal Information: Environmental and Molecular Mutagenesis, Journal Name: Environmental and Molecular Mutagenesis Journal Issue: Suppl.23 Vol. 23; ISSN 0893-6692; ISSN EMMUEG
• Country of Publication: United States
• Language: English

Related Subjects

56 BIOLOGY AND MEDICINE
APPLIED STUDIES
BIOLOGICAL REPAIR
CHO CELLS
DOSE-RESPONSE RELATIONSHIPS
GAMMA RADIATION
GENETIC RADIATION EFFECTS
MESSENGER-RNA