Measurements of Rate of Transcription in Isolated Nuclei by Nuclear "Run-Off' Assay

Abstract

AbstractUnlike gene expression in prokaryotic cells, which is primarily under transcriptional control, gene expression in eukaryotic cells is subject to both transcriptional and posttranscriptional controls. Since transcription and translation in eukaryotic cells are separated topographically, the regulation of mRNA metabolism can occur at multiple sites, within nuclei and in the cytoplasm. Nevertheless, transcription remains a critical locus of control of eukaryotic gene expression. Transcriptional regulation affects cellular mRNA abundance by affecting rates of tran- scription. Another important control mechanism that can affect mRNA abundance is the rate of mRNA decay. Thus, half-life of mRNA repre- sents a balance between rates of transcription and intracellular degrada- tion, e.g., an increase in the abundance of mRNA could result from decreased mRNA degradation, increased mRNA synthesis, or both.KeywordsSucrose SolutionSulfonyl ChlorideGuanidinium ThiocyanateLabel IncorporationGeiger CounterThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.