Method and compositions for detecting botulinum neurotoxin
AssigneeWisconsin Alumni Research Foundation
InventorsEdwin R. Chapman, Min Dong
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AssigneeWisconsin Alumni Research Foundation
InventorsEdwin R. Chapman, Min Dong
Estimating the term…
A molecular construct capable of fluorescent resonance energy transfer (FRET), comprising a linker peptide, a donor fluorophore moiety and an acceptor fluorophore moiety, wherein the linker peptide is a substrate of a botulinum neurotoxin selected from the group consisting of synaptobrevin, syntaxin and SNAP-25, or a fragment thereof capable being cleaved by the botulinum neurotoxin, and separates the donor and acceptor fluorophores by a distance of not more than 10 nm, and wherein emission spectrum of the donor fluorophore moiety overlaps with the excitation spectrum of the acceptor fluorophore moiety; or wherein the emission spectra of the fluorophores are detectably different. Also provided are isolated nucleic acid expressing the construct, kits comprising said construct and cell lines comprising said nucleic acid. Further provided are methods of detecting a BoNT using the above described construct via FRET, and methods for detecting a BoNT using surface plasmon resonance imaging.
An isolated polynucleotide molecule encoding a molecular construct, wherein the construct is capable of anchoring to a plasma membrane or a vesicular membrane of a cell, the construct comprising (1) a linker peptide, (2) a FRET pair comprising a donor fluorophore moiety and an acceptor fluorophore moiety, and (3) a membrane-anchoring domain that directs the anchoring of the construct to the plasma or vesicular membrane of the cell, wherein the linker peptide comprises a cleavage recognition site of a botulinum neurotoxin selected from the group consisting of synaptobrevin, syntaxin and SNAP-25, and wherein the membrane-anchoring domain comprises a fragment that contains the palmitoylation site (residues 83-120) of SEQ ID NO:
Dependencies are inferred from the claim text.