|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
16
|
|
pubmed:dateCreated |
1991-10-4
|
|
pubmed:abstractText |
We have established a sensitive, monoclonal antibody (Mab)-based procedure permitting the selective enrichment of sequences containing the miscoding alkylation product O6-ethylguanine (O6-EtGua) from mammalian DNA. H5 rat hepatoma cells were reacted with the N-nitroso carcinogen N-ethyl-N-nitrosourea in vitro, to give overall levels of greater than or equal to 25 O6-EtGua residues per diploid genome (corresponding to O6-EtGua/guanine molar ratios of greater than or equal to 10(-8). For analysis, enzymatically restricted DNA from these cells is incubated with an antibody specific for O6-ethyl-2'-deoxyguanosine, the resulting Mab-DNA complexes are separated from (O6-EtGua)-free fragments by filtration through a nitrocellulose (NC) membrane, and the DNA is recovered from the filter-bound complexes quantitatively. The efficiency of Mab binding to DNA fragments containing O6-EtGua is constant over a range of O6-EtGua/guanine molar ratios between 10(-5) and 10(-8). (O6-EtGua)-containing restriction fragments encompassing known gene sequences (e.g., the immunoglobulin E heavy chain gene of H5 rat hepatoma cells used as a model in this study) are subsequently amplified by PCR and quantified by slot-blot hybridisation. The content and distribution of a specific carcinogen-DNA adduct in defined sequences of genomic DNA can thus be analyzed as well as the kinetics of intragenomic (toposelective) repair of any DNA lesion for which a suitable Mab is available.
|
|
pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2202730,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2203764,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2263639,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2295129,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2387020,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2407530,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2554145,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2785688,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-2999980,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3046938,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3329097,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3349485,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3415677,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3466163,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3525535,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3664636,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3698104,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3742723,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3786142,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3838150,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3855560,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3923365,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-3948328,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-4085447,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-545720,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6086939,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6251549,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6324102,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6370677,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6704400,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6733565,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-6746711,
http://linkedlifedata.com/resource/pubmed/commentcorrection/1886770-7471102
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Aug
|
|
pubmed:issn |
0305-1048
|
|
pubmed:author |
|
|
pubmed:issnType |
Print
|
|
pubmed:day |
25
|
|
pubmed:volume |
19
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
4467-72
|
|
pubmed:dateRevised |
2009-11-18
|
|
pubmed:meshHeading |
pubmed-meshheading:1886770-Animals,
pubmed-meshheading:1886770-Antibodies, Monoclonal,
pubmed-meshheading:1886770-Base Sequence,
pubmed-meshheading:1886770-DNA,
pubmed-meshheading:1886770-DNA Damage,
pubmed-meshheading:1886770-Deoxyguanosine,
pubmed-meshheading:1886770-Ethylnitrosourea,
pubmed-meshheading:1886770-Kinetics,
pubmed-meshheading:1886770-Molecular Sequence Data,
pubmed-meshheading:1886770-Polymerase Chain Reaction,
pubmed-meshheading:1886770-Rats,
pubmed-meshheading:1886770-Tumor Cells, Cultured
|
|
pubmed:year |
1991
|
|
pubmed:articleTitle |
Monoclonal antibody-based, selective isolation of DNA fragments containing an alkylated base to be quantified in defined gene sequences.
|
|
pubmed:affiliation |
Institute of Cell Biology (Cancer Research), University of Essen Medical School, FRG.
|
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|
