In vivo biotinylation of proteins using a single vector system
Author Information
Author(s): Predonzani Andrea, Arnoldi Francesca, López-Requena Alejandro, Burrone Oscar R
Primary Institution: International Centre for Genetic Engineering and Biotechnology
Hypothesis
Can a single bigenic plasmid system be used for site-specific biotinylation of proteins in vivo?
Conclusion
The study demonstrates that a single bigenic plasmid can effectively induce site-specific biotinylation of both secretory and membrane-bound proteins.
Supporting Evidence
- A single bigenic plasmid was developed for the co-expression of a protein tagged with BAP and an engineered BirA enzyme.
- Three different recombinant antibodies were biotinylated and maintained their binding properties.
- The method allows for the purification of biotinylated proteins from culture supernatants.
Takeaway
Scientists created a new way to attach biotin to proteins in living cells using just one piece of DNA, making it easier to study and use these proteins.
Methodology
The study involved constructing a single bigenic plasmid for co-expressing a biotin acceptor peptide and the BirA enzyme in mammalian cells.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website