Novel pristinamycin-responsive expression systems for plant cells

Research output: Contribution to journalArticle

Researchers

  • Alexander Frey

  • Markus Rimann
  • James E. Bailey
  • Pauli T. Kallio
  • Charles J. Thompson
  • Martin Fussenegger

Research units

  • Swiss Federal Institute of Technology Zurich

Abstract

Novel gene regulation systems were designed for plant cells responsive to the streptogramin antibiotic pristinamycin. The pristinamycin-repressible plant gene regulation concept (PIPpOFF) is based on a transcriptional activator (PIT) which consists of the Pip protein, the repressor of the pristinamycin resistance operon of Streptomyces coelicolor, fused to the VP16 transactivation domain of the Herpes simplex virus. PIT mediates pristinamycin-repressible activation of a synthetic plant promoter (PpPIR) in tobacco cells consisting of a nine Pip-binding site-containing artificial operator (PIR3) placed upstream of a TATA-box derived from the cauliflower mosaic virus 35S promoter (PcaMV35S). Pristinamycin interferes with induction by negatively regulating the DNA-binding capacity of the Pip moiety of PIT. A second, streptogramin-inducible plant gene regulation system (PIPpON) was constructed by combining Pip expression with a plant-specific pristinamycin-inducible promoter (PpPIRON). PpPIRON consists of a PIR3 module cloned downstream of the strong constitutive plant promoter PCaMV35S. AS in the native Streptomyces configuration, Pip binds to its cognate sequence within PpPIRON in the absence of regulating antibiotic and silences the chimeric plant promoter. Upon addition of pristinamycin, Pip is released from the PIR3 operator and full PCaMV35S-driven expression of desired plant genes is induced. The PIPpOFF and PIPpON systems performed well in Nicotiana tabacum suspension cultures and promise to provide an attractive extension of existing plant gene regulation technology for basic plant research or biopharmaceutical manufacturing using plant tissue culture.

Details

Original languageEnglish
Pages (from-to)154-163
Number of pages10
JournalBiotechnology and Bioengineering
Volume74
Issue number2
Publication statusPublished - 16 May 2001
MoE publication typeA1 Journal article-refereed

    Research areas

  • Pip, Plant gene regulation, Pristinamycin, Streptogramin, Streptomyces, Tobacco

ID: 4506552