title: Transcription in Archaea: in vitro transcription assays for mjRNAP.
abstract: The fully recombinant Methanocaldococcus jannaschii RNA polymerase allows for a detailed dissection of the different stages of the transcription. In the previous chapter, we discussed how to purify the different components of the M. jannaschii transcription system, the RNA polymerase subunits, and general transcription factors and how to assemble a functional M. jannaschii enzyme. Standard in vitro transcription assays can be used to examine the different stages of transcription. In this chapter, we describe how some of these assays have been optimized for M. jannaschii RNA polymerase, which transcribes at much higher temperatures than many other transcription complexes.
authors: Institute of Structural and Molecular Biology, University College London, Darwin Building, Gower Street, London, WC1E 6BT, 020 7679 3899, UK, k.smollett@ucl.ac.uk.
authors: Katherine
authors: K
authors: Smollett
authors: Fabian
authors: F
authors: Blombach
authors: Finn
authors: F
authors: Werner
mesh_terms: D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D066298:In Vitro Techniques / Q000379:methods*; D063747:Methanocaldococcus / Q000201:enzymology* / Q000502:physiology*; D021122:Protein Subunits / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D013696:Temperature; D014158:Transcription, Genetic / Q000235:genetics / Q000502:physiology*
chemical_list: D021122:Protein Subunits; D011994:Recombinant Proteins; D012321:DNA-Directed RNA Polymerases