Ribonucleic acid synthesis by chromatin isolated from Phaseolus aureus Roxb.

Chromatin was isolated from the hypocotyls of Phaseolus aureus Roxb. and assayed for endogenous RNA polymerase (EC 2.7.7.6) activity in vitro. The molecular size of the RNA product, measured by polyacrylamide gel electrophoresis, was found to be much smaller than that known to be synthesised in vivo and was affected by the assay temperature. Although conventional enzyme assays provided no evidence for the presence of ribonuclease in chromatin, a more sensitive technique revealed sufficient ribonuclease activity to degrade high-molecular weight RNA to smaller fragments. The inclusion of unlabelled exogenous RNA in the media for chromatin preparation and RNA polymerase assay substantially increased the molecular-weight of the RNA products synthesised in vitro.Abbreviations TCA trichloroacetic acid - UTP uridine-5-triphosphate - UMP uridine-5-monophosphate - SOS sodium dodecyl sulphate