A comparative analysis of recombinant protein expression in different biofactories: Bacteria, insect cells and plant systems

Journal of Visualized Experiments

Volumn 2015, Issue 97, 2015, Pages

  Gecchele, Elisa ^a   Merlin, Matilde ^a   Brozzetti, Annalisa ^b   Falorni, Alberto ^b   Pezzotti, Mario ^a   Avesani, Linda ^a  

^a UNIVERSITY OF VERONA   (Italy)

^b UNIVERSITY OF PERUGIA   (Italy)

Author keywords

Baculovirus insect cells; Issue 97; Nicotiana benthamiana; Nicotiana tabacum; Plant biology; Plant biotechnology; Recombinant protein; Stable expression; Transgenic plant; Transient expression

Indexed keywords

GLUTAMATE DECARBOXYLASE; GLUTAMATE DECARBOXYLASE 2; RECOMBINANT PROTEIN;

ANIMAL; BACULOVIRIDAE; BIOSYNTHESIS; BIOTECHNOLOGY; COMPARATIVE STUDY; ENZYMOLOGY; ESCHERICHIA COLI; GENETICS; HUMAN; METABOLISM; PROCEDURES; SF9 CELL LINE; SPODOPTERA; TOBACCO; TRANSGENIC PLANT; VIROLOGY;

ANIMALS; BACULOVIRIDAE; BIOTECHNOLOGY; ESCHERICHIA COLI; GLUTAMATE DECARBOXYLASE; HUMANS; PLANTS, GENETICALLY MODIFIED; RECOMBINANT PROTEINS; SF9 CELLS; SPODOPTERA; TOBACCO;

EID: 84941236302     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/52459     Document Type: Article

References (15)

1. Hampe, C. S., Hammerle, L. P., Falorni, A., Robertson, J., Lernmark, A. Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding. FEBS Lett. 488, (3), 185-189 (2001).
2. Avesani, L., et al. Recombinant human GAD65 accumulates to high levels in transgenic tobacco plants when expressed as an enzymatically inactive mutant. Plant Biotechnol. J. 9, (8), 862-872 (2010).
3. Sambrook, J., et al. Molecular Cloning: A laboratory manual. Second Edition. Cold Spring Harbor Laboratory Press New York (1989).
4. Avesani, L., et al. Comparative analysis of different biofactories for the production of a major diabetes autoantigen. Transgenic Res. 23, 281-291 (2014).
5. Marillonnet, S., Giritch, A., Gils, M., Kandzia, R., Klimyuk, V., Gleba, Y. In planta engineering of viral RNA replicons: efficient assembly by recombination of DNA modules delivered by Agrobacterium. Proc. Natl. Acad. Sci. (USA). 101, (18), 6852-6857 (2004).
6. Gleba, Y., Klimyuk, V., Marillonnet, S. Viral vectors for the expression of proteins in plants. Curr. Opin. Biotechnol. 18, 134-141 (2007).
7. Engler, C., Kandzia, R., Marillonnet, S. A one pot, one step, precision cloning method with high throughput capability. PLoS One. 3, (11), (2008).
8. Xu, R., Li, Q. Q. Protocol: streamline cloning of genes into binary vectors in Agrobacterium via the Gateway TOPO vector system. Plant Methods. 4, (4), 1-7 (2008).
9. Fairbanks, G., Steck, T. L., Wallach, D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 10, (13), 2606-2617 (1971).
10. Falorni, A., et al. Radioimmunoassay detects the frequent occurrence of autoantibodies to the Mr 65,000 isoform of glutamic acid decarboxylase in Japanese insulin-dependent diabetes. Autoimmunity. 19, 113-125 (1994).
11. Hunt, I. From gene to protein: a review of new and enabling technologies for multi-parallel protein expression. Protein Expr. Purif. 40, (1), 1-22 (2005).
12. Arzola, L., et al. Transient co-expression of post-transcriptional silencing suppressor for increased in planta expression of a recombinant anthrax receptor fusion protein. Int. J. Mol. Sci. 12, (8), 4975-4990 (2011).
13. Merlin, M., Gecchele, E., Capaldi, S., Pezzotti, M., Avesani, L. Comparative evaluation of recombinant protein production in different biofactories: the green perspective. Biomed. Res. Int. 2014, 136419 (2014).
14. Avesani, L., et al. Improved in planta expression of the human islet autoantigen glutamic acid decarboxylase (GAD65). Transgenic Res. 12, (2), 203-212 (2003).
15. Leuzinger, K., et al. Efficient agroinfiltration of Plants for high-level transient expression of recombinant proteins. J Vis Exp. (77), (2013).