FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology, genetic and protein engineering and is intended for the production of genetically fused hybrid proteins. Recombinant DNA plasmids pFLAG-sc14D5a-NLuc-r1 and pFLAG-NLuc-r2-sc14D5a containing a unique gene of a single-stranded mouse antibody directed against tick-borne encephalitis virus protein E and genes of NLuc-r1 and NLuc-r2 luciferase fragments are proposed. A hybrid protein is proposed, expressed in Escherichia coli Rosetta-gami2 cells transformed by the mentioned plasmid DNA, followed by isolation from the cytoplasmic fraction and purification by metal-chelate chromatography of hybrid proteins sc14D5a-NLuc-r1 and NLuc-r2-sc14D5a.
EFFECT: present invention enables to obtain two hybrid proteins with the ability to bind tick-borne encephalitis virus proteins E with the complementation of NanoLuc luciferase fragments and the occurrence of bioluminescent activity in the presence of a substrate - celenterazine, as a potentially suitable highly sensitive system for detecting tick-borne encephalitis virus in biological samples by single-phase bioluminescent analysis.
3 cl, 9 dwg, 6 ex
| Title |
Year |
Author |
Number |
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2012 |
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2022 |
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1997 |
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