¹Federal State Budgetary Institution «Research Institute of Experimental Medicine», North Western Branch of the RAMSs, Saint Petersburg, Russian Federation
²2Hannover Medical School, Hannover, Germany
*Corresponding author: Prof Michael M. Shavlovsky, PhD, ScD, Head of Human Molecular Genetics Laboratory, Department of Molecular Genetics, Federal State Budgetary Institution «Research Institute of Experimental Medicine», North Western Branch of the RAM S, 12, Acad. Pavlov str., 197376, Saint Petersburg, Russian Federation. Tel: 7-812-2343356 Fax: 7-812-3205849. E-Mail:mmsch@rambler.ru
We investigated the possibility of obtaining native recombinant amyloidogenic proteins by creating genetic constructs encoding fusion proteins of target proteins with Super Folder Green Fluorescent Protein (sfGFP). In this study, we show that the structures, containing the sfGFP gene, provide a synthesis, within a bacterial system, of fusion proteins with minimal formation of inclusion bodies. Constructs containing genes of the target proteins in the 3'-terminal region of the sfGFP gene followed by a polynucleotide sequence, which allows for affinity purification fusion proteins, are optimal. Heating bacterial cultures before the induction of the expression of recombinant genes in 42°C for 30 min (heat shock) was found to increase the output of the desired products, thus practically avoiding the formation of insoluble aggregates.
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