Microbial Physiology
Short Communication
Expression and Purification of Heterologous Proteins in Plant Tissue Using a Geminivirus Vector SystemHefferon K.L.a · Kipp P.c · Moon Y.S.baCornell Research Foundation and bDepartment of Horticulture, Plant Sciences Building, Cornell University, Ithaca,N.Y., and cLexicon Genetics Incorporated, TheWoodlands,Tex., USA
|
|
Log in to MyKarger to check if you already have access to this content.
KAB
Buy a Karger Article Bundle (KAB) and profit from a discount!
If you would like to redeem your KAB credit, please log in.
Save over 20% compared to the individual article price.
Article / Publication Details
Published online: July 22, 2004
Issue release date: July 2004
Number of Print Pages: 6
Number of Figures: 3
Number of Tables: 0
ISSN: 2673-1665 (Print)
eISSN: 2673-1673 (Online)
For additional information: https://www.karger.com/MIP
Abstract
In the past, plant molecular biologists have relied on Escherichia coli, baculovirus and other expression systems to produce plant proteins to quantities sufficient for biochemical analysis. However, such expression systems often result in the production of proteins which possess improper posttranslational modifications. Here, we present a plant virus-based expression system superior to those currently available. We demonstrate that bean yellow dwarf geminivirus (BeYDV) replicates and expresses foreign proteins at high levels in tobacco, Arabidopsis, and other dicotyledonous plants, making it more universal than plant RNA viruses with restricted host ranges which are currently used as expression systems. The DNA-based nature of the BeYDV genome renders it stable for the incorporation of large plant open reading frames, and gives it an advantage over other plant virus-based expression systems which possess insert size restrictions. Using this expression system, the rapid accumulation of a novel Arabidopsis-derived mitogen-activated protein kinase to levels sufficient for standard biochemical analysis is demonstrated.
© 2004 S. Karger AG, Basel
Related Articles:
References
- Dowd C, Bekes F: Large-scale expression and purification of high-molecular-weight glutenin subunits. Protein Expr Purif 2002;25/1:97–104.
- Edens L, Heslinga L, Klok R, Ledeboer AM, Mas J, Toonen MY, Visser C, Verrips CT: Cloning of cDNA encoding the sweet-tasting plant protein thaumatin and its expression in Escherichia coli. Gene 1982;18/1:1–12.
- Ferrini JB, Martin M, Taupiac MP, Beaumelle B: Expression of functional ricin B chain using the baculovirus system. Eur J Biochem 1995;233:772–777.
- Galili G: Heterologous expression of a wheat high molecular weight glutenin gene in Escherichia coli. Proc Natl Acad Sci USA 1989;86:7756–7760.
- Hayes RJ, MacDonald H, Coutts RHA, Buck KW: Priming of complementary DNA synthesis in vitro by small DNA molecules tightly bound to virion DNA of wheat dwarf virus. J Gen Virol 1988;69:1345–1350.
- Hefferon KL, Dugdale BG: Independent expression of Rep and RepA and their roles in regulating bean yellow dwarf virus replication. J Gen Virol 2003;84:3465–3472.
- Hefferon KL, Xu J, AbouHaidar MG: Identification and in vivo expression of a prokaryotic-like ribosome recognition sequence located upstream of the coat protein gene of potato virus X. Arch Virol 2000;145:945–956.
- Henderson J, Atkibnson AE, Lazarus CM, Hawes CR, Napier RM, Macdonald H, King LA: Stable expression of maize auxin-binding protein in insect cell lines. FEBS Lett 1995;371/3:293–296.
- Innes REW: Mapping out the roles of MAP kinases in plant defense. Trends Plant Sci 2001;6:392–394.
-
Jarvis DL: Baculovirus expression vectors; in Miller LK (ed): The Baculoviruses. New York, Plenum Press, 1997.
- Kammann M, Matzeit V, Schell J, Walden R, Gronenborn B: Geminivirus-based shuttle vectors capable of replication in Escherichia coli and monocotyledonous plant cells. Gene 1991;104:247–252.
- Kunze R, Fusswinkel H, Feldmar S: Expression of plant proteins in baculoviral and bacterial systems. Methods Cell Biol 1995;50:461–479.
- Liu L, Davies JW, Stanley J: Mutational analysis of bean yellow dwarf virus, a geminivirus of the genus mastrevirus that is adapted to dicotyledonous plants. J Gen Virol 1998;79:2265–2274.
- Liu L, van Tonder T, Pietersen G, Davies JW, Stanley J: Molecular characterization of a subgroup I geminivirus from a legume in South Africa. J Gen Virol 1997;78:2113–2117.
- MAPK Group: Mitogen-activated protein kinase cascades in plants: A new nomenclature. Trends Plant Sci 2002;7:301–308.
- Mockaitis K, Howell SH: Auxin induces mitogenic activated protein kinase (MAPK) activation in roots of Arabidopsis seedlings. Plant J 2000;24:785–796.
- Muren E, Rask L: Processing in vitro of pronapin, the 2S storage-protein precursor of Brassica napus produced in a baculovirus expression system. Planta 1996;200:373–379.
- Palmer KE, Rybicki EP: The molecular biology of mastreviruses. Adv Virus Res 1998;50:183–234.
-
Paszty C, Lurquin PF: Improved plant protoplast plating/selection technique for quantitation of transformation frequencies. Biotechniques 1987;5:716–718.
External Resources
- Porta C, Lomonosoff GP: Use of viral replicons for the expression of genes in plants. Mol Biotechnol 1996;5/3:209–221.
-
Sambrook J, Fritschef EF, Maniatis T: Molecular Cloning: A Laboratory Manual, ed 2. New York, Cold Spring Harbor Laboratory Press, 1989.
- Suzuki K: MAP kinase cascades in elicitor signal transduction. J Plant Res 2002;115/3:237–244.
-
Timmermans MCP, Das OP, Messing J: Geminiviruses and their uses as extrachromosomal replicons. Annu Rev Plant Physiol Plant Mol Biol 1994;45:79–112.
External Resources
- Tozzini AC, Ek B, Palva T, Hopp HE: Potato virus X coat protein: A glycoprotein. Virology 1994;202:651–658.
- Woolston CJ, Reynolds BV, Stacey NJ, Mullineaux PM: Replication of wheat dwarf virus DNA in protoplasts and analysis of coat protein mutants in protoplasts and plants. Nucleic Acids Res 1989;17:6029–6041.
- Wrzaczek M, Hirt H: Plant MAP kinase pathways: How many and what for? Biol Cell 2001;93/1–2:81–87.
- Zwerger K, Hirt H: Recent advances in plant MAP kinase signaling. Biol Chem 2001;382:1123–1131.
Article / Publication Details
Published online: July 22, 2004
Issue release date: July 2004
Number of Print Pages: 6
Number of Figures: 3
Number of Tables: 0
ISSN: 2673-1665 (Print)
eISSN: 2673-1673 (Online)
For additional information: https://www.karger.com/MIP
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Get Permission