Isolation of genomic DNA fragments corresponding to genes modulated in vivo by a transcription factor.
A new methodology for the identification of genes modulated by transcription factors in vivo is described. Mouse genomic DNA fragments bound by the thyroid hormone receptor (T3R) were selected and amplified in vitro. Subsequent hybridisation with biotinylated cDNA allowed the selection of those DNA fragments containing binding sites for T3R that corresponded to transcribed DNA. Expression analysis of the corresponding genes showed that more than 80% are indeed modulated by thyroid hormones in vivo in the liver. Together with the presence of consensus binding sites for T3R this result suggests that the selected DNA fragments may contain T3R transcriptional regulatory elements. This method, extensive to other ligand-modulated transcription factors, might be useful to all transcription factors with slight modifications.
Caubín
J
J
Departamento de Investigación, Pharmacia Antibióticos Farma S.A., Madrid, Spain.
Iglesias
T
T
Bernal
J
J
Muñoz
A
A
Márquez
G
G
Barbero
J L
JL
Zaballos
A
A
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001710:Biotin; D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D015342:DNA Probes; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D006980:Hyperthyroidism / Q000378:metabolism; D007037:Hypothyroidism / Q000378:metabolism; D007163:Immunosorbent Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011988:Receptors, Thyroid Hormone / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000494:pharmacology*
D015342:DNA Probes; D011988:Receptors, Thyroid Hormone; D014157:Transcription Factors; D001710:Biotin; D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases