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EXPRESSION OF MUTANT FORM OF EGFP IN VIRAL VECTOR
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| EXPRESSION OF MUTANT FORM OF EGFP IN VIRAL VECTOR
Gritsenko D., Deryabina N. Institute of Plant biology and Biotechnology al-Farabi Kazakh National University d.kopytina@gmail.com GFP (Green fluorescence protein) is a secondary emitter in the bioluminescent system of jellyfish (Aequorea victoria) and is capable of fluorescence in the green range when exposed to blue light emitted by aequorin at a wavelength of 469 nm. The use of GFP as a reporter protein provides tremendous opportunities in the study of cell organization; processes of vital activity of cells, tissues; transport of molecules inside the cell; localization of certain proteins, protein complexes; to track the sorting of proteins within the cell; gene expression, protein- protein interactions; development of viral and bacterial infections in cells, as well as the development of universal protein expression systems for medicine and farming. Currently, there are many mutant forms of GFP, which are divided into 7 classes depending on the quality of the chromophore. Second class includes proteins with phenolate anions in the chromophore, e.g. EGFP, Emerald and so on. EGFP may include the following variants: S65 mutations; F64L, S65T; F64L, S65T, V163A. The absorption peak is 488-489 nm and the emission peak is 509-511 nm. In our work, the EGFP protein (F64L, S65T) with the mutation at position 90 (Pro → Ser) was examined. The replacement of the hydrophobic amino acid by hydrophilic in the alpha chain, on which the chromophore is localized. The investigation of the mutation using computer programs showed no effect on the formation of the secondary structure of the protein. Expression of mutant EGFP in plants using a viral vector based on the genome of grapevine virus A was confirmed by confocal microscopy and immunoblot analysis. Scientific adviser: PhD Galiakparov N. жүктеу/скачать 2,79 Mb. Достарыңызбен бөлісу:
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