Unpublished
New England Biolabs (NEB) has engineered a SNAP-CBD (chitin-binding domain) fusion protein that can be used in the preparation of chitin-binding probes. The labeling of SNAP-CBD with an appropriate SNAP-tag substrate is a highly efficient reaction that allows for consistent and effective labeling with good yields. The labeling procedure itself is robust and should be straightforward to conduct in most labs. Furthermore, the availability of SNAP-tag substrates bearing many different fluorophores increases the versatility of the SNAP-CBD protein by enabling the production of a wide variety of chitin-binding probes.
NEB developed a fusion construct comprised of Bacillus circulans CBD and a SNAP-tag protein to take advantage of the SNAP-tag labeling system, which allows for convenient and precise labeling with near-quantitative efficiency. With this strategy, one single fluorophore molecule reacts with the SNAP-tag portion of a fusion construct, thus avoiding the under- or over-labeling that is often observed with other chemical labeling techniques. Internal studies at NEB have shown that fluorescently-labeled SNAP-CBD is equally specific and efficient in detecting chitin in C. elegans as the earlier CBD-MBP probes. In addition, the availability of different SNAP-reactive fluorescent substrates as well as non-fluorescent substrates expands the utility of CBD in research. Many SNAP-tag-reactive substrates are commercially available from NEB.
Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 104586 | pYZ205 | Express SNAP-6His-CBD |