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Protein Subcellular Localization in Plant
Creative Biogene offers protein subcellular localization service to help customers understand the cellular functions of target protein in an organism. We utilize three different plant subcellular localization systems, including protoplasts, tobacco leaf disc, onion epidermal cells to visualize the location of target genes in plant subcellular structures. Our professional scientists will flexibly handle the project to accelerate your research on gene function and protein interaction.
Accurate protein subcellular localization detection is necessary for the targeting and functional characterization of the proteins of interest. There are a variety of methods for determining subcellular localization, including immunofluorescent labeling, fluorescent protein fusion, co-localization with marker enzyme. The most common is to use fluorescence microscopy to compare the localization of your protein of interest with known markers.
The process of fluorescent protein fusion method
First, the ORF sequence of target gene is fused with a fluorescent protein (such as GFP, BFP, CFP, YFP, OFP, mCherry, mKATE) and cloned into a suitable expression construct. Then, expression vector of the fusion gene and organelle marker are co-transformed into plant cells (protoplast, tobacco leaf disc, onion epidermal cell). Finally, observing the localization of the fluorescent protein by fluorescence microscope or laser confocal microscope to determine the subcellular localization of the target protein.
Creative Biogene develops several localization systems to help clients visualize the location of target protein in living plant cell.
- Protoplasts localization system
Plant protoplasts can be used as a cell-based experimental model to introduce macromolecules such as DNA, RNA and proteins using PEG-mediated transformation. Plant protoplast is also regarded as the most precision model for subcellular localization, due to all organelles can be observed clearly using fluorescence microscope. Creative Biogene establishes protoplast isolation and transformation system in various species, to meet your subject requirements.
Figure 1. Subcellular localization analysis in Arabidopsis protoplasts
- Tobacco leaf disc localization system
Tobacco leaf disc system is the most convenient and cost-effective cell-based model to analysis the localization and interaction of target protein for all plants species. Creative Biogene utilizes Agrobacterium-mediated transient transformation technology to monitor the subcellular localization for gene of interest.
Figure 2. Subcellular localization analysis in tobacco leaf.
- Onion epidermal cell localization system
Onion epidermal cell is a suitable cell-based subcellular localization model for gene which localized in cell wall. Onion plasmolysis can be used to distinguish the target gene location in cell wall and plasma membrane, which cannot achieve in two other systems.
Advantages
- Various optional localization system: Arabidopsis/rice/maize/tobacco/wheat/tobacco protoplast, tobacco leaf, onion epidermal cell.
- Multiple organelle marker: chloroplast, cytoplasm, endoplasmic reticulum, mitochondrion, nucleus, peroxisome, plastid, vacuole and plasma membrane.
- Various fluorescent protein: GFP, BFP, CFP, YFP, OFP, mCherry, mKATE
- One-stop solution: from gene synthesis/cloning, construction, transient transformation to fluorescence microscopy imaging.
Creative Biogene offers one-stop solution of subcellular localization analysis in plant. Our highly experienced staffs will cooperate with you to identify a sound and cost-effective solution to meet your needs.
References:
- S., E., (2005). Iron-sulfur cluster biogenesis in chloroplasts. involvement of the scaffold protein cpisca. Plant Physiology.
- Xi, N., (2018). A rice cpyc-type glutaredoxin osgrx20 in protection against bacterial blight, methyl viologen and salt stresses. Frontiers in Plant Science, 9, 111.
- Gong, Q. Q., (2015). Ostctp, encoding a translationally controlled tumor protein, plays an important role in mercury tolerance in rice. Bmc Plant Biology, 15(1), 123.