OBTAINING VECTOR CONSTRUCTS FOR THE EXPRESSION OF THE FIRST SUBUNIT OF HEMAGGLUTININ INFLUENZA VIRU TYPE
At the moment, there is considerable concern about the spread in the world of epizootics of the avian influenza virus. Influenza virus has the highest genetic variability and the likelihood of new strains that can create large epidemics. The evolution of the influenza virus proceeds very quickly, therefore, the paramount task of the researchers is antigenic mapping of hemagglutinin subtypes, as well as identifying the characteristics of the antigenic structure. X-ray crystallography is a commonly used method for determining the three-dimensional structure of a protein.
The purpose of these studies was to obtain vector constructs for the expression of the protein of the first subunit of influenza A virus for further expression in the bacterial system.
As a result of the studies, specific primers for amplification of the first subunit of hemagglutinin gene were synthesized. When designing oligonucleotides for the greatest specificity, all the criteria for primers for PCR were taken into account. The restriction endonucleases NcoI and XhoI were chosen for primers, the sites of which are absent in the sequence of the gene itself and are present in the multiclonal region of the plasmid. Also, in the course of the studies, a genetic construct was obtained for the expression of the first subunit of hemagglutinin protein in E. coli cells. The obtained recombinant protein will be used for further work on crystallography and three-dimensional modeling of the protein.