Genetically Modified Coelenterazine-Dependent Luciferases as Reporters for In Vitro Assay

Alexander N. Kudryavtsev, Vasilisa V. Krasitskaya, Eugenia E. Bashmakova, Ludmila A. Frank
2017 Журнал Сибирского федерального университета: Серия Биология  
Among the most important tasks of modern biotechnology is the development of new analytical techniques suitable for highly sensitive and specific detection of diagnostically important targets rapidly expanding their range with the progress in biomedical science. In this view, particular attention is paid to elaboration of methods based on photosensitive technologies (fluorescent, chemiluminescent, etc.). The studies on bioluminescent systems of various luminous organisms are being carried out
more » ... being carried out in the laboratory of photobiology of the Institute of Biophysics SB RAS for more than 30 years. The genes of many light-emitting proteins, photoproteins and luciferases, were cloned. The recombinant protein analogues were obtained, their spatial structures were determined and the mechanisms of functioning were proposed. With site-directed mutagenesis applied, the luciferase variants with unique beneficial properties were created. Highly sensitive analytical systems for diagnosing a number of biologically active compounds -hormones, oncogenes, infectious agents, etc. were developed on the basis of bioluminescent reactions of these proteins. The paper presents the recent years' results of these studies. These are the method of simultaneous detection of two targets in one sample on the basis of color variants of Ca 2+ -regulated photoprotein obelin and the method for rapid and sensitive detection of tick-borne encephalitis virus based on the unique biospecific fusion protein of Renilla luciferase. The research results testify that the proposed analytical systems are competitive and applicable in modern medical laboratory.
doi:10.17516/1997-1389-0023 fatcat:7vuf4w5ywvbzbg2yei6b3lbk5a