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    Zpusob identifikace farmakologicky ucinných látekzaložený na in vitro transkripci
    11.
    发明专利
    Zpusob identifikace farmakologicky ucinných látekzaložený na in vitro transkripci 未知

    公开(公告)号：CZ298459B6

    公开(公告)日：2007-10-10

    申请号：CZ72598

    申请日：1998-03-10

    Applicant: HOECHST AG

    Inventor： KIRSCHBAUM BERND ,  STAHL WILHELM ,  WINKLER IRVIN ,  MEISTERERNST MICHAEL

    IPC: C12N15/09 ,  C12N15/10 ,  C12N15/11 ,  C12N15/63 ,  C12P19/34 ,  C12Q1/68

    Abstract: Popisuje se automatizovatelný zpusob transkripce in vitro urcený pro identifikaci farmakologicky úcinných látek zahrnující kroky, kdy se a) poskytne DNA templát, který obsahuje cílovou DNA sekvenci pod kontrolou alespon jednoho genového regulacníhoelementu, b) s DNA templátem smíchá jaderný extrakt a alespon jeden znacený nukleotid, c) s DNA templátem se smíchá kandidátní farmakologicky úcinná látka, d) prípadne se po in vitro transkripci DNA templátu z reakcní smesi odstraní proteiny, e) znacený transkript se naváže na pevný nosic, f) odstraní se nadbytek znacených nukleotidu, a g) stanovíse množství znaceného transkriptu vzhledem ke kontrolnímu vzorku, pricemž DNA templát obsahuje jedinecná rozpoznávací místa pro restrikcní endonukleázy PstI, EcoRI, SacI, KpnI, SacII, BamHI, SwaI, cást plazmidu pUC19, pet vazebných míst pro Ga14 protein z kvasinek, "TATA" BOX Z REPTORU V.beta.8.1 humánních T lymfocytu mezi restrikcními místy SacIIa BamHI, iniciacní oblast adenovirového hlavního pozdního promotoru mezi restrikcními míst BamHI a SwaI, a sekvenci bez G-nukleotidu délky 800 páru bází.

    In-vitro transcription processes for screening natural products and other chemical substances
    13.
    发明专利
    In-vitro transcription processes for screening natural products and other chemical substances 未知

    公开(公告)号：AU5836098A

    公开(公告)日：1998-09-17

    申请号：AU5836098

    申请日：1998-03-11

    Applicant: HOECHST AG

    Inventor： KIRSCHBAUM BERND ,  STAHL WILHELM ,  WINKLER IRVIN ,  MEISTERERNST MICHAEL

    IPC: C12N15/09 ,  C12N15/63 ,  C12P19/34 ,  C12Q1/68 ,  C12N15/00

    Abstract: Process for transcription of a DNA template in vitro in a cell-free system, where the template comprises a sequence to be transcribed under the control of at least one regulatory element, is characterised in that (a) an enriched and optionally purified extract of cell nuclei and at least one labelled nucleotide are used for the transcription, where the extract can optionally be supplemented or partly or completely replaced by transcription factors and/or cofactors, (b) proteins present in the reaction mixture at the end of transcription are optionally separated and/or degraded, (c) the labelled transcript is bound to a solid support, (d) excess labelled nucleotide is removed, and (e) the amount of labelled transcript is determined. Also claimed is a DNA template as above in which the sequence to be transcribed does not contain at least one nucleobase selected from G, C or T, where the G-, C- or T-free sequence has a length of more than 400 nucleotides.

    METHOD OF CELL-FREE IN VITRO TRANSCRIBING A DNA MATRIX, DNA MATRIX A SUCH AND APPLICATION OF THAT METHOD AND SUCH DNA MATRIX
    14.
    发明专利
    METHOD OF CELL-FREE IN VITRO TRANSCRIBING A DNA MATRIX, DNA MATRIX A SUCH AND APPLICATION OF THAT METHOD AND SUCH DNA MATRIX 未知

    公开(公告)号：PL325343A1

    公开(公告)日：1998-09-14

    申请号：PL32534398

    申请日：1998-03-12

    Applicant: HOECHST AG

    Inventor： KIRSCHBAUM BERND ,  STAHL WILHELM ,  WINKLER IRVIN ,  MEISTERERNST MICHAEL

    IPC: C12N15/09 ,  C12N15/63 ,  C12P19/34 ,  C12Q1/68 ,  C12N15/11 ,  C07H21/04

    Abstract: Process for transcription of a DNA template in vitro in a cell-free system, where the template comprises a sequence to be transcribed under the control of at least one regulatory element, is characterised in that (a) an enriched and optionally purified extract of cell nuclei and at least one labelled nucleotide are used for the transcription, where the extract can optionally be supplemented or partly or completely replaced by transcription factors and/or cofactors, (b) proteins present in the reaction mixture at the end of transcription are optionally separated and/or degraded, (c) the labelled transcript is bound to a solid support, (d) excess labelled nucleotide is removed, and (e) the amount of labelled transcript is determined. Also claimed is a DNA template as above in which the sequence to be transcribed does not contain at least one nucleobase selected from G, C or T, where the G-, C- or T-free sequence has a length of more than 400 nucleotides.

    IN-VITRO TRANSCRIPTION PROCESSES FOR SCREENING NATURAL PRODUCTS AND OTHER CHEMICAL SUBSTANCES
    19.
    发明专利
    IN-VITRO TRANSCRIPTION PROCESSES FOR SCREENING NATURAL PRODUCTS AND OTHER CHEMICAL SUBSTANCES 未知

    公开(公告)号：CA2231745C

    公开(公告)日：2009-11-03

    申请号：CA2231745

    申请日：1998-03-11

    Applicant: HOECHST AG

    Inventor： KIRSCHBAUM BERND ,  STAHL WILHELM ,  WINKLER IRVIN ,  MEISTERERNST MICHAEL

    IPC: C12N15/09 ,  C12N15/63 ,  C12N15/11 ,  C12P19/34 ,  C12Q1/68

    Abstract: 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 an d 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, purifie d 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 labele d 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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