Modeling of the induced expression for high-level production of a foreign protein by recombinant E. coli under the control of the T7 phage promoter

TOKSOY ÖNER E., Kirdar B., Onsan Z., Ulgen K.

PROCESS BIOCHEMISTRY, cilt.39, sa.3, ss.315-323, 2003 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 39 Sayı: 3
  • Basım Tarihi: 2003
  • Doi Numarası: 10.1016/s0032-9592(03)00077-3
  • Dergi Adı: PROCESS BIOCHEMISTRY
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.315-323
  • Anahtar Kelimeler: Escherichia coli, structured modeling, compartment, Taq I, induced expression, T7 phage promoter, TAQI RESTRICTION-ENDONUCLEASE, ESCHERICHIA-COLI, STRUCTURED MODEL, RNA-POLYMERASE, CULTURES, FERMENTATION, SYSTEM, GROWTH
  • Marmara Üniversitesi Adresli: Evet

Özet

A structured four-compartment mathematical model is proposed for describing the behaviour of three expression systems. In system [R], Taq I endonuclease gene was expressed under the control of T7 RNA polymerase promoter in recombinant Escherichia coli cells. Methylase protection was provided either by cloning the Taq I methylase gene on to a second plasmid (system [R, M]) or by constructing a recombinant plasmid harbouring both genes (system [R+M]). Considering the promoter strength, the inducer effect is included in the rate equation for foreign protein production. A separate set of parameters was estimated for simulations of the uninduced and induced phases (dual-parameter set) of systems [R] and [R, M]. A single parameter set describing both phases was also determined. Simulation results indicate that the dual-parameter approach is more successful in predicting the experimental observations. Model validation conducted on system [R+M] using parameter sets of system [R, M] revealed good agreement. (C) 2003 Elsevier Science Ltd. All rights reserved.