Coordinate expression of antibody subunit genes yields high levels of functional antibodies in roots of transgenic tobacco

Autor: Fred A. van Engelen, Alexander Schouten, Jos W. Molthoff, Jan Roosien, Jesús Salinas, Wim G. Dirkse, Arjen Schots, Jaap Bakker, Fred J. Gommers, Maarten A. Jongsma, Dirk Bosch, Willem J. Stiekema
Jazyk: angličtina
Rok vydání: 1994
Předmět:
medicine.drug_class
Molecular Sequence Data
protein assembly
Enzyme-Linked Immunosorbent Assay
Plant Science
Chimeric gene
Monoclonal antibody
Immunoglobulin light chain
Plant Roots
Antigen-Antibody Reactions
Transformation
Genetic
Antigen
Tobacco
Genetics
medicine
Amino Acid Sequence
Cloning
Molecular
Promoter Regions
Genetic
Gene
Laboratorium voor Nematologie
Antibodies
Fungal
Antibody
coordinated expression
Plant Diseases
genetic engineering
biology
Base Sequence
Antibodies
Monoclonal
Promoter
General Medicine
Plants
Genetically Modified
root
Molecular biology
secretion
Plants
Toxic
Mycoses
biology.protein
Plantibody
Immunoglobulin Light Chains
EPS
Laboratory of Nematology
Immunoglobulin Heavy Chains
Agronomy and Crop Science
Carboxylic Ester Hydrolases
Protein Processing
Post-Translational
Zdroj: Plant Molecular Biology 26 (1994)
Plant Molecular Biology, 26, 1701-1710
ISSN: 0167-4412
Popis: To explore the feasibility of employing antibodies to obtain disease resistance against plant root pathogens, we have studied the expression of genes encoding antibodies in roots of transgenic plants. A model monoclonal antibody was used that binds to a fungal cutinase. Heavy and light chain cDNAs were amplified by PCR, fused to a signal sequence for secretion and cloned behind CaMV 35S and TR2′ promoters in a single T-DNA. The chimeric genes were cloned both in tandem and in a divergent orientation. The roots of tobacco plants transformed with these constructs produced antibodies that were able to bind antigen in an ELISA. Immunoblotting showed assembly to a full-size antibody. In addition, a F(ab′)2-like fragment was observed, which is probably formed by proteolytic processing. Both antibody species were properly targeted to the apoplast, but the full-size antibody was partially retained by the wall of suspension cells. The construct with divergent promoters showed a better performance than the construct with promoters in tandem. It directed the accumulation of functional antibodies to a maximum of 1.1% of total soluble protein, with half of the plants having levels higher than 0.35%. The high efficiency of this construct probably results from coordinated and balanced expression of light and heavy chain genes, as evidenced by RNA blot hybridization.
Databáze: OpenAIRE
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