Lentil root protoplasts: a transient expression system suitable for coelectroporation of monoclonal antibodies and plasmid molecules

Mauro Maccarrone, Gerrit A. Veldink, Alessandro Finazzi Agrò, Johannes F G Vliegenthart

Research output: Contribution to journalArticle

Abstract

Protoplasts were isolated from lentil (Lens culinaris) roots and their suitability as a transient expression system was investigated. After transfecting the protoplasts with the β-glucuronidase (GUS) gene by either electroporation or polyethylene glycol (PEG), the specific activity of the reporter enzyme and the cell viability were determined. Electroporation was more effective than PEG treatment as transfection procedure and its efficiency was affected by the plasmid length. The feasibility of electro-transferring at the same time (coelectroporation) inhibitory anti-lipoxygenase monoclonal antibodies and the GUS-carrying plasmid pBI 221 was investigated as well. The amount of transferred immunoglobulins was quantitated by ELISA and the inhibitory ability of monoclonal antibodies on the intracellular target enzyme was determined. Evidence is presented for the successful coelectroporation of immunoglobulins and plasmid DNA into lentil protoplasts, the two types of macromolecules acting independently of each other in the recipient cells.

Original languageEnglish
Pages (from-to)136-142
Number of pages7
JournalBBA - General Subjects
Volume1243
Issue number1
DOIs
Publication statusPublished - Jan 18 1995

Keywords

  • Coelectroporation
  • L. culinaris
  • Lipoxygenase
  • Monoclonal antibody
  • Protoplast
  • Transient expression

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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