Patent ID: 6015682
Filing Date: 2000-01-18
Classification: C07K,C12N,G01N

Abstract:
A method of identifying a substance that modifies gene transcription, comprising the steps of:a) combining:(i) DNA to be transcribed;(ii) transcription factor a, or a homolog thereof;(iii) TATA-binding protein;(iv) a test substance to be assessed for its ability to modify gene transcription; and(v) a purified RNA polymerase II holoenzyme preparation comprising general transcription factors b, g, and e and at least eight SRB proteins, wherein said SRB polypeptides are selected from the group consisting of SRB2, SRB4, SRB5, SRB6, SRB7, SRB8, SRB9, SRB10, and SRB11, and wherein at least one of said SRB polypeptides binds specifically to a polyclonal antibody raised against S. cerevisiae SRB2, SRB4, SRB5, SRB6, SRB7, SRB8, SRB9, or human SRB7, thereby producing a test mixture;(b) maintaining the test mixture of step a) under conditions sufficient for transcription of the DNA; and(c) determining the extent to which DNA transcription occurs in the test mixture and comparing the result of such determination with the extent to which DNA transcription occurs in a DNA transcription reaction according to steps a) and b) in the absence of said test substance; said test substance being one that modifies gene transcription when DNA transcription occurs to a lesser or greater extent in said test mixture than in said DNA transcription reaction performed in the absence of test substance.