FIELD: biotechnology.
SUBSTANCE: group of inventions relates to biotechnology and is intended to assay the enzyme activity of botulinum neurotoxin. Provided is a reporting comprising: a membrane anchoring domain, a cleavage site and a reporter domain comprising two identical reporter peptides. Enzyme activity at the cleavage site releases the reporter region into the cytosol, where degradation events occur under cell protease action. Also disclosed is a method for assaying the activity of botulinum neurotoxin, in which a cell comprising an endogenous protease and expressing a reporter construct is provided. Said cell contacts with a sample suspected of including the enzyme activity of botulinum neurotoxin. Decrease in the detectable signal at a first wavelength is observed when the sample includes the enzyme activity of botulinum neurotoxin.
EFFECT: inventions provide a rapid, sensitive and accurate assay for enzyme activities associated with botulinum toxins, permit the use of the same construct for assays aimed at more than one analyte with enzyme activity, providing improved access to more recognition sites within the cleavage domain of said construct.
24 cl, 24 dwg, 1 ex
