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., ÖNSAN Z. İ., ÜLGEN Ş. K.

Process Biochemistry, cilt.39, sa.3, ss.315-323, 2003 (SCI-Expanded, Scopus)

  • 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: Compartment, Escherichia coli, Induced expression, Structured modeling, T7 phage promoter, Taq I
  • Boğaziçi Üniversitesi Adresli: Evet

Özet

A structured four-compartment mathematical model is proposed for describing the behaviour of three expression systems. In system [R], Tag 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. © 2003 Elsevier Science Ltd. All rights reserved.