Selection of primers for the obtaining of subunits of the avian influenza virus hemagglutinin gene

Abstract

Influenza A virus, in contrast to influenza viruses B and C, affects many species of mammals and birds and is very variable. Particularly variable are the surface glycoproteins of the virion, haemagglutinin (HA) and neu-raminidase (NA). The purpose of this work is to select primers and optimize the conditions for obtaining the nucleotide sequence of the hemagglutinin gene of the avian influenza virus and its subunits by the polymerase chain reaction method for further determination of its three-dimensional structure. The paper presents the results of work on the selection and synthesis of primers, including sites for endonuclease cleavage of the restriction enzymes NcoI and XhoI, flanking sections of the NA gene. These primers are characterized by rather high sensitivity, specificity and reproducibility of the results. The synthesized specific primers were used to amplify the HA gene and its specific regions, and the PCR conditions were optimized. The results of the conducted studies are expedient for using in the works on obtaining the recombinant hemagglutinin protein of the avian influenza virus, which will be used todetermine its three-dimensional structure.