In-vitro transcription processes for screening natural...

Details In-vitro transcription processes for screening natural... In-vitro transcription processes for screening natural...

C - Chemistry, Metallurgy

12

N

C12N 15/63 (2006.01) C12N 15/11 (2006.01) C12P 19/34 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2231745

The invention relates to an in-vitro process for analyzing transcription of viral and cellular genes which can be automated and which is suitable for efficient and economical bulk screening with the aim of finding specific chemical lead structures which have a selective effect on gene activity. The process for the cell-free in-vitro transcription of a DNA template comprising a DNA sequence to be transcribed which is under the control of at least one gene-regulatory element comprises a) using, for the transcription, a concentrated and, if appropriate, purified extract from cell nuclei which, if appropriate, can be complemented, or partially or fully replaced, by transcription factors and/or cofactors, and at least one labeled nucleotide, b) if appropriate isolating and/or degrading the proteins in the reaction mixture after transcription, c) binding the labeled transcript to a solid matrix, d) removing the excess labeled nucleotides and e) determining the amount of labeled transcript.

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