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Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins
Published on: July 23, 2013
49.4K
Systems and strategies for plant protein expression
Anthony J Zmuda1, Thomas D Niehaus1
1Department of Plant and Microbial Biology, University of Minnesota, St. Paul, MN, United States.
Methods in Enzymology
|January 29, 2023
Summary
This study details methods for expressing and purifying plant enzymes, NAD(P)HX dehydratase (NAXD) and NAD(P)HX epimerase (NAXE), aiding in understanding plant metabolism and cofactor repair. These strategies can be applied to other plant proteins.
Area of Science:
- Plant biochemistry
- Enzymology
- Molecular biology
Background:
- A significant portion of plant genes encode uncharacterized enzymes, hindering a complete understanding of plant metabolism.
- Functional characterization of these enzymes is crucial for advancing plant science.
Purpose of the Study:
- To describe practical strategies for the heterologous expression and purification of Arabidopsis thaliana and Zea mays NAD(P)HX dehydratase (NAXD) and NAD(P)HX epimerase (NAXE).
- To provide protocols applicable to a broader range of plant proteins.
Main Methods:
- Transit peptide prediction and construct design for recombinant protein expression.
- Heterologous expression in Escherichia coli.
- Purification protocols for target enzymes.
Main Results:
- Successful expression and purification protocols were established for NAXD and NAXE from *Arabidopsis thaliana* and *Zea mays*.
- The developed strategies are broadly applicable to other plant proteins.
Conclusions:
- The described methods facilitate the functional and structural characterization of uncharacterized plant enzymes.
- This work contributes to deciphering plant metabolism and the repair mechanisms of NAD(P)H cofactors.
Keywords:
Arabidopsis thalianaEscherichia coliHeterologous expressionMetabolite damageMetabolite repairNAD(P)HXPlant enzyme expressionPlant metabolismRecombinant protein expressionTransit peptide predictionMore Related Videos
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