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Nucleic Acids Research, 1994, Vol. 22, No. 20 4132-4138
© 1994

METHODS

Isolation of genomic DNA fragments corresponding to genes modulated in vivo by a transcription factor

Julio Caubín, Teresa lglesias$, Juan Bernal1, Alberto Muñoz1, Gabriel Márquez, José Luis Barbero and Angel Zaballos*

Departamento de Investigatión, Pharmacia Antibiéoticos Farma S.A. Antonio López 109, 28026 Madrid 1Instituto de Investigaciones Biomédicas (CSIC) Arturo Duperier 4, 28029 Madrid, Spain

*To whom correspondence should be addressed

Received July 5, 1994. Revised September 9, 1994. Accepted September 9, 1994.

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 ligandmodulated transcription factors, might be useful to all transcription factors with slight modifications.

$Present address, Instituto de Investigaciones Biometiicas (CSIC), Arturo Duperier 4, 28029 Madrid


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