Prokaryotic Expression of the Influenza A (H5N1) Neuraminidase

Abstract

Background and purpose: Highly Pathogenic Influenza A(H5N1) viruses cause vast economic losses throughout the world. They may circulate in animals and be able to spread from human to human. Therefore, launching diagnostic tests are highly essential to control the influenza infection. Studies on the amino acid sequence of the neuraminidase (NA) protein of influenza viruses revealed that NA is the most immunogenic protein in naïve animals which can simply stimulate the humoral immune system well. Materials and methods: Influenza virus NA gene (A/Indonesia/5/2005(H5N1) was cloned into pET21a and expressed in host E. coli (BL21) strains. Then, the expression level of NA protein was optimized for different IPTG inductor concentrations and times. Results: Findings showed the integrity of pET21a-NA construct. Emerging bands with the expected molecular weight (38KDal) on SDS-PAGE and WB analysis confirmed the successful expression of target protein in E.coli BL21 strain. In silico analysis showed integrity of major epitopes in the structure of fused version of NA produced in this work. Conclusion: The new recombinant NA has the potential to be used directly in serological tests. It could be also used in polyclonal antibody preparation which is employed as an essential material in western blot analyses and other immunological and serological studies, such as ELISA, immunocytochemistry, and immunohistochemistry.

Keywords

• neuraminidase
• prokaryote system
• diagnostic tests
• influenza a (h5n1) virus