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Cloning and Expression of Recombinant Nucleoprotein of Influenza H1N1

Somaie Tavakoli, Mojgan Bandehpour, Zeinab Soleimanifar, Mehdi Goudarzi, Bahram Kazemi
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Abstract

Background: Influenza virus is the major cause of lower respiratory tract illnesses on the worldwide. Vaccination can be an effective tool to prevent its outbreak. Highly conserved viral nucleoprotein is an effective vaccine candidate to provide heterosubtypic immunity, offering resistance against various influenza virus strains.

Materials and Methods: In present research NP gene was inserted in pET-22b expression vector. New construct (pET-22b/NP) was transformed into E. coli BL21 (DE3) strain and the expression of nucleoprotein was induced by IPTG. It was analyzed by SDS-PAGE and confirmed by Western blotting.

Results: Western blotting confirmed the expression and production of recombinant Influenza nucleoprotein.

Conclusion: These results suggest that the codon-optimized influenza A virus NP gene can be efficiently expressed in E. coli.


Keywords

Influenza virus, Cloning, Expression, Nucleoprotein gene

References

Dongming Z. A Universal Influenza A Vaccine Based on Adenovirus Expressing Matrix-2 Ectodomain and Nucleoprotein Protects Mice From Lethal Challenge. Mol Ther. 2010; 1812: 2182–9.

Horimoto T, KawaokaY. Influenza: lessons from past pandemics, warnings from current incidents. Nat Rev Microbiol. 2005;3:591–600.

Nicholson KG, Wood JM, Zambon M. Influenza. Lancet. 2003;362:1733–45.

Epstein SL, Kong W, Misplon JA, Loa CY, Tumpey TM, Gary LX et al. Protection against multiple influenza A subtypes by vaccination with highly conserved nucleoprotein. Vaccine. 2005; 23:5404–10.

Ohba K, Yoshid S, Dewan Z, Shimura H, Sakamaki N, Takeshita F, et al. Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross-reactivity. Vaccine. 2007;25:4291–300.

Huang B, Wang W, Li R, Wang X, Jiang T, Qi X, Influenza A virus nucleoprotein derived from Escherichia coli or recombinant vaccinia (Tiantan) virus elicits robust cross-protection in mice. Virol J. 2012;9:322.

Kazemi B, Seyed N, Bandehpour M, Sharifnia Z, Pakzad P. Cloning Expression and Purification of Truncated Chlamydia Trachomatis Outer Membrane Protein 2 (Omp2) and its Application in an ELISA Assay. Iran J Immunol. 2008;5(3):148-55.

Bandehpour M, Seyed N, Shadnoush M, Pakzad P, Kazemi B.Using recombinant Chlamydia Major Outer Membrane Protein (MOMP) in ELISA diagnostic Kit. Iran J BIOTECH. 2006;4(4):239-44.

Kenji Ohba. Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross-reactivity. Vaccine. 2007;25:4291–300.

Fields virology. Orthomyxoviruses. 5th ed. Lippincott Williams & Wilkins, Philadelphia, PA. 2007;2:1691–740.

Price GE, Soboleski MR, Chia-Yun Lo, Misplon JA, Quirion MR, Houser KV. Single-Dose Mucosal Immunization with a Candidate Universal Influenza Vaccine Provides Rapid Protection from Virulent H5N1, H3N2 and H1N1 Viruses. PLoS One. 2010;5(10):13162.

Salahuddin P, Khan AU. Structure function studies on different structural domains of nucleoprotein of H1N1 subtype. Bioinformatics. 2010;5(1):28–30.

Portela A, Digard P. The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. J General Virol. 2002;83:723-34.




DOI: https://doi.org/10.22037/nbm.v3i2.7137