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»
ABSTRACT. The article describes the construction of vectors and the carrying out of bacterial expression of recombinant FSH of bovine animals (bFSH) for the production of milligram quantities of its protein subunits. Glycosilated protein bovine follicle stimulating hormone (bFSH) is one of the gonadotropin family, playing important role in reproduction and can be obtained in different systems for heterological expression to preserve its native structure. E. coli was chosen as a host for production of milligram quantities of this protein, because even nonglycosilated bacterial-derived form of the recombinant bFSH to a certain extent preserves biological activity. The major part of alfa and beta subunits of FSH protein is synthesized in form of inclusion bodies, regardless of type of the vector (pET-28a(+) and pET40b(+)) and temperature of expression used (16°C and 37°C). Biosynthesis in the form of inclusion bodies requires that they be solubilized in denatured conditions with following protein refolding into the right shape to function. The washing of inclusion bodies with following ammonium sulphate fractionation leads to 80% pure protein. Simultaneous expression of separate subunits in the same host cell is possible and has some advantages. Non-glycosylated bFSH, as well as its individual subunits, can be used in further studies, including research of their native conformation and biological action, with the aim of developing new effective approaches for improving reproductive technologies in livestock.
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