Abstract
This study provides a mathematical model of T7 RNA polymerase (T7 RNAP)
kinetics under in vitro conditions targeted at application of this model
to simulation of dynamic transcription performance. A functional
dependence of transcript synthesis rate is derived based on: (a)
essential reactant concentrations, including T7 RNAP and its promoter,
substrate nucleotides, and the inhibitory byproduct inorganic
pyrophosphate; (b) a distinction among vector characteristics such as
recognition sequences regulating transcription initiation and
termination, respectively; and (c) specific properties of the nucleotide
sequence including both transcript length and nucleotide composition.
Inactivation kinetics showed a half-life of T7 RNAP activity of 50 min
under the conditions applied in vitro using the isolated enzyme. Model
parameters and their precision are estimated using dynamic simulation
and nonlinear regression analysis. The particular novelty of this model
is its capability to incorporate linear genomic sequence information for
simulation of nonlinear in vitro transcription kinetics. (C) 2001 John
Wiley & Sons, Inc.
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