Journal of Vaccines & Vaccination

Prokaryotic expression of influenza conserved domain chimer protein (HA2-NP) as a universal vaccine candidate

13^th Annual Congress on Vaccines, Therapeutics & Travel Medicine: Influenza & Infectious diseases

December 01-02, 2016 Atlanta, USA

Fatemeh Morshedi, Farahmand B, Nazeri E, Fotouhi F, Shokouhi H and Saleh M

Pasteur Institute of Iran, Iran
Islamic Azad University, Iran

Posters & Accepted Abstracts: J Vaccines Vaccin

Abstract:

Influenza, a respiratory pathogen, causes high degree of mortality and morbidity during seasonal epidemics and sporicidal pandemics. By selecting conserved antigenic proteins, e.g. hemagglutinin small subunit (HA2) and nucleoprotein (NP), we would be able to develop a vaccine based on a fusion protein causing both cellular and humoral responses which are the most challenging aspects in designing a universal vaccine. For this purpose, primers for the antigenic part of NP were designed using bioinformatic tools. The desired product was multiplied via polymerase chain reaction (PCR) method using the designed primers which was then penetrated into T vector, followed by insertion into pET28a vector in order to construct pET28a/NP. Both pET28a/NP and pET28a/ HA2, the latter was previously generated in our lab, were digested with the same restriction enzymes HindIII/Xho1 and therefore NP was inserted to downstream region of HA2 in pET28a/HA2 construct. The generated pET28a/HA2-NP was transformed into E. coli (BL21(DE3)) using chemical competent (CaCl2) method. The expression was induced by isopropylbeta-D-thiogalactopyranoside (IPTG) in concentration of 1 mM in 37°C and optimized in 28°C. The results were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by western blotting. The results showed that the antigenic segment of NP was successfully cloned into pET28a/HA2. The 36 kDa protein band of HA2-NP was observed on SDS-PAGE and western blotting. Unlike current available vaccines in which some allergic reactions are observed, our chimer protein is continual and safe, stimulating both cellular and humoral immunity systems. Our construct could potentially provide a basis for a universal vaccine candidate.

Biography :

Email: fatemeh_morshedi1368@yahoo.com