Agrobacterium-Mediated Transient Gene Expression Optimized for the Bioenergy Crop Camelina sativa

被引:0
|
作者
Kumar, Pawan [1 ,2 ]
Banday, Zeeshan Z. [1 ]
Riley, John L. [1 ]
Greenberg, Jean T. [1 ]
机构
[1] Univ Chicago, Dept Mol Genet & Cell Biol, Chicago, IL 60637 USA
[2] Univ Chicago, Dept Ecol & Evolut, Chicago, IL USA
来源
BIO-PROTOCOL | 2024年 / 14卷 / 07期
关键词
ARABIDOPSIS;
D O I
10.21769/BioProtoc.4964
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Camelina sativa, a Brassicaceae family crop, is used for fodder, human food, and biofuels. Its relatively high resistance to abiotic and biotic stresses, as well as being a climate-resilient oilseed crop, has contributed to its popularity. Camelina 's seed yield and oil contents have been improved using various technologies like RNAi and CRISPR/Cas9 genome editing. A stable transformation system for protein localization and other cell autonomous investigations, on the other hand, is tedious and time consuming. This study describes a transient gene expression protocol for Camelina sativa cultivar DH55 leaves using Agrobacterium strain C58C1. The method is suitable for subcellular protein localization and colocalization studies and can be used with both constitutive and chemically induced genes. We report the subcellular localization of the N -terminal ER membrane signal anchor region (1-32 aa) of the At3G28580 gene-encoded protein from Arabidopsis in intact leaves and the expression and localization of other known organelle markers. This method offers a fast and convenient way to study proteins in the commercially important Camelina crop system.
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页数:10
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