FIELD: biotechnology.
SUBSTANCE: to implement this method, nucleosomes are collected from purified histones on DNA matrices, then ligated with RNA polymerase, and the molecular target PARP1 is introduced into the resulting complex, followed by the transcription buffer, as well as the test chemical compounds. The said transcription buffer contains deoxynucleotide triphosphates (dNTPs) and labeled α-32F-GTF. Next, transcription and evaluation of transcription products are carried out by their separation with electrophoresis. The conclusion about the presence of PARP1 inhibitory activity in the test compound is made by presence bands at 15, 45 and in the run-off region on the electrophoregram, showing transcription pausing, as shown in Fig. 1, the said activity rating being carried out with a reference value of PARP1 with NAD.
EFFECT: invention allows to increase the specificity and selectivity of a nucleosome-based molecule selection system.
19 cl, 2 dwg, 7 ex
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Authors
Dates
2017-12-21—Published
2015-12-07—Filed