Всероссийский научно-исследовательский институт физиологии, биохимии и питания животных – филиал Федерального государственного бюджетного научного учреждения «Федеральный научный центр животноводства – ВИЖ имени академика Л.К. Эрнста»
Bacterial producers of recombinant proteins derived from hantavirus Dobrava Gc protein were engineered. They exhibit normal viability and high yield of the product sufficient for its preparative purification. The obtained result experimentally confirmed a theoretical model of Gc domain organization including a hypothesis that a putative fusion loop should be eliminated from the structure as far as it demonstrates a high motility and is prone to interactions with membranes. A system for in vivo quantification of the recombinant protein yield in a soluble form using color fluorescent proteins has been established. pYNC-Dob construct protein (full-length Gc protein depleted of fusion loop, transmembrane and cytoplasmic domain fused with 6His-EYFP carrier) is accumulated in cytoplasm of the producer strains with 93 mg/l yield. Soluble protein shares app. 10% of this amount. The product conserves fluorescence ability in PAAG under “semi-denaturing” conditions of the sample preparing.
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