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The peptide sequence GLNDIFEAQKIEWHE is known as the AviTag, a 15-amino-acid peptide widely used in molecular biology for site-specific biotinylation of proteins. This tag is recognized by the biotin ligase enzyme BirA from Escherichia coli, which catalyzes the covalent attachment of biotin to the lysine residue within the AviTag sequence.
Functional Overview
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Biotinylation Site: The AviTag contains a specific lysine residue that serves as the attachment point for biotin, facilitated by BirA.
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Specificity: BirA-mediated biotinylation is highly specific, ensuring that biotin is attached only to the AviTag sequence, minimizing off-target modifications.
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Applications:
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Protein Purification: Biotinylated proteins can be efficiently purified using streptavidin or avidin affinity systems.
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Protein Immobilization: Biotinylated proteins can be immobilized on streptavidin-coated surfaces for various assays.
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Protein Detection: Biotinylated proteins can be detected using streptavidin-conjugated probes in techniques like Western blotting or ELISA.
Research and Development
The AviTag has been incorporated into various recombinant protein constructs to facilitate studies in protein-protein interactions, enzymatic activity assays, and structural biology. Its small size minimizes interference with the native function of the fused protein, making it a versatile tool in protein engineering.
The GLNDIFEAQKIEWHE sequence, or AviTag, is a powerful tool in molecular biology that enables precise biotinylation of proteins, facilitating their purification, immobilization, and detection. Its specificity and efficiency have made it a standard in various biochemical and biotechnological applications.
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