TY - JOUR
T1 - Lentil root protoplasts
T2 - a transient expression system suitable for coelectroporation of monoclonal antibodies and plasmid molecules
AU - Maccarrone, Mauro
AU - Veldink, Gerrit A.
AU - Finazzi Agrò, Alessandro
AU - Vliegenthart, Johannes F G
PY - 1995/1/18
Y1 - 1995/1/18
N2 - 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.
AB - 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.
KW - Coelectroporation
KW - L. culinaris
KW - Lipoxygenase
KW - Monoclonal antibody
KW - Protoplast
KW - Transient expression
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U2 - 10.1016/0304-4165(94)00124-G
DO - 10.1016/0304-4165(94)00124-G
M3 - Article
C2 - 7827102
AN - SCOPUS:0028812315
VL - 1243
SP - 136
EP - 142
JO - Biochimica et Biophysica Acta - General Subjects
JF - Biochimica et Biophysica Acta - General Subjects
SN - 0304-4165
IS - 1
ER -