This control plasmid contains the gene encoding the Histone H2B protein cloned upstream of the SNAPf coding sequence in pSNAPf, as a fusion to the N- terminus of the SNAP-tag. Histone H2B is a member of the core histones that package DNA in the nucleus. The H2B-SNAPf fusion protein gives nuclear fluorescence when labeled with SNAP-Cell® substrates. The full sequence and map for pSNAPf-H2B can be downloaded.
The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). SNAP-tag substrates are derivatives of benzylguanines and benzylchloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.
pSNAPf -H2B contains an improved version of SNAP-tag, termed SNAPf. SNAPf displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.
There are two steps to using this system: sub cloning and expression of the protein of interest as a SNAPf fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Expression of the SNAPf -H2B fusion protein is described in this document. The labeling of fusion proteins with SNAP-tag substrates is described in the instructions supplied with SNAP-tag substrates.
DNASU is a central repository for plasmid clones and collections that may also be helpful.
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