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Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation.

Authors
Jung, Y; Rhee, Y; Auh, CK; Shim, H; Choi, JJ; Kwon, ST; Yang, JS; Kim, D; Kwon, MH; Kim, YS; Lee, S
Citation
Plant cell reports, 28(10):1593-1602, 2009
Journal Title
Plant cell reports
ISSN
0721-77141432-203X
Abstract
We developed an asexual reproductive plant, Kalanchoe pinnata, as a new bioreactor for plant-based molecular farming using a newly developed transformation method. Leaf crenate margins were pin-pricked to infect the plant with the Agrobacterium strain LBA4404 and vacuum infiltration was also applied to introduce the target gene into the plants. Subsequently, the young mother leaf produced new clones at the leaf crenate margins without the need for time- and labor-consuming tissue culture procedures. The average transformation rates were approximately 77 and 84% for pin-prickling and vacuum-infiltration methods, respectively. To functionally characterize an introduced target protein, a nucleic acid hydrolyzing recombinant 3D8 scFv was selected and the plant based 3D8 scFv proteins were purified and analyzed. Based on abzyme analysis, the purified protein expressed with this system had catalytic activity and exhibited all of properties of the protein produced in an E. coli system. This result suggested that vegetatively reproductive K. pinnata can be a novel and potent bioreactor for bio-pharmaceutical proteins.
MeSH terms
Gene Transfer Techniques*Genetic VectorsImmunoglobulin Variable Region/biosynthesis*Immunoglobulin Variable Region/isolation & purificationKalanchoe/geneticsKalanchoe/immunologyKalanchoe/metabolism*Plants, Genetically Modified/geneticsPlants, Genetically Modified/immunologyPlants, Genetically Modified/metabolismRNA, Plant/geneticsRhizobium/geneticsTransformation, Genetic
DOI
10.1007/s00299-009-0758-3
PMID
19688214
Appears in Collections:
Journal Papers > School of Medicine / Graduate School of Medicine > Microbiology
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