BIOSENSOR FOR DETERMINING INTRACELLULAR TEMPERATURE AND VISCOSITY BASED ON GENETICALLY MODIFIED PHOTOACTIVE ORANGE CAROTENOID PROTEIN Russian patent published in 2020 - IPC C07K14/00 C07K19/00 C12Q1/00 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, particularly to biosensors for measuring intracellular temperature and viscosity. Genetically modified water-soluble orange carotenoid protein (OCP) protein with one cysteine in position 245 contains as a chromophore carotenoid molecule (3'-hydroxyxyinonone or canthaxanthin) and has photoconversion, that is when absorbing blue-green light, the protein changes from orange to red. Genetically modified OCP protein is conjugated with fluorescent TMR label through cysteine in position 245 of protein. After illumination with blue-green light, OCP protein turns into red, thus increasing the distance between the fluorescent label and the carotenoid molecule, disrupting the energy migration process, and the label begins to fluoresce. This process depends on temperature and viscosity of environment. Accuracy of measuring temperature in solution is 0.2 °C ± 0.05 °C.

EFFECT: due to the presence of a carotenoid in the protein portion, the biosensor has low cytotoxicity and is able to deactivate the ROS which can be formed during measurement.

6 cl, 5 dwg, 1 ex