Specificity of chromatin transcription in vitro. Asymmetric transcription of the globin gene by Escherichia coli RNA polymerase.
The transcription of globin genes in mouse foetal liver chromatin and nuclei by exogenous Escherichia coli RNA polymerase is prone to artifacts due to RNA-dependent transcription of endogenous mRNA sequences. This is particularly evident when Mn2+ is used as divalent cation in the RNA transcription reaction. However, substitution or supplementation with Mg2+ eliminates this artifact and gives essentially asymmetric DNA-dependent transcription by the polymerase. In this paper we discuss a number of general criteria which can be applied to test the validity of specific gene transcription in vitro.
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism; D008274:Magnesium / Q000502:physiology; D051379:Mice; D014158:Transcription, Genetic
D002843:Chromatin; D005914:Globins; D012321:DNA-Directed RNA Polymerases; D008274:Magnesium