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Engineering, Cloning and Expression of Interleukin 2–Com1 Chimera with Aim of Recombinant Subunit Vaccine Production Against Coxiella burnetii

  • Amin Jaydari
  • Peyman Esmaeili Fard Barzegar
  • Ali Forouharmehr
  • Ali Kakanezhadifard
  • Narges Nazifi
Article
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Abstract

Query fever is an important disease caused by Coxiella burnetii, therefore vaccination against this disease is so crucial. Com1 is one the most important immunogenic proteins of C. burnetii which can be appropriate candidate to design a subunit vaccine. It seems, fusion of this protein with interleukin 2 as a molecular adjuvant can promote its efficacy. To do current study, first, Com1 and interleukin 2 genes were individually amplified by PCR, then these PCR products were applied to fuse interleukin 2 with Com1 using splice overlap extension PCR. The product of splice overlap extension PCR was cloned in pTZ57R/T vector by T/A cloning strategy, after digestion, interleukin 2–Com1 gene was sub-cloned in pET-22b(+) by T4 DNA ligase enzyme. Ligation product was applied to express in BL21 (DE3) strain of Escherichia coli. Expressed interleukin 2–Com1 protein was purified by Nik–NTA affinity column and then confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. The results of electrophoresis on agarose 1% gel revealed that, PCR of target genes and splice overlap extension PCR were successfully performed. The results of electrophoresis on 12% sodium dodecyl sulfate polyacrylamide gel confirmed expression and purification of interleukin 2–Com1 protein. Finally, the results of western blotting shown, purified protein with a molecular size of 43.5 kDa belonged IL2–Com1 chimera. The results of present study shown, BL21 (DE3) is an appropriate host to express interleukin 2–Com1 protein. It seems, this chimera can be introduced as a potent candidate to fight with Query fever.

Keywords

Coxiella burnetii Com1 Subunit vaccine Interleukin 2 Protein expression 

Notes

Acknowledgements

The authors would like to thank research deputy of Loretan University due to its support from this project.

References

  1. Amet N, Lee H-F, Shen W-C (2009) Insertion of the designed helical linker led to increased expression of tf-based fusion proteins. Pharm Res 26(3):523CrossRefPubMedPubMedCentralGoogle Scholar
  2. Bai Y, Shen W-C (2006) Improving the oral efficacy of recombinant granulocyte colony-stimulating factor and transferrin fusion protein by spacer optimization. Pharm Res 23(9):2116–2121CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bai Y, Ann DK, Shen W-C (2005) Recombinant granulocyte colony-stimulating factor-transferrin fusion protein as an oral myelopoietic agent. Proc Natl Acad Sci 2(20):7292–7296CrossRefGoogle Scholar
  4. Chen X, Zaro JL, Shen W-C (2013) Fusion protein linkers: property, design and functionality. Adv Drug Deliv Rev 65(10):1357–1369CrossRefPubMedPubMedCentralGoogle Scholar
  5. Forouharmehr A, Nassiri M, Ghovvati S, Javadmanesh A (2018) Evaluation of different signal peptides for secretory production of recombinant bovine pancreatic ribonuclease A in Gram negative bacterial system: an in silico study. Curr Proteomics 15(1):24–33CrossRefGoogle Scholar
  6. Gerlach C, Škultéty Ľ, Henning K, Neubauer H, Mertens K (2017) Coxiella burnetii immunogenic proteins as a basis for new Q fever diagnostic and vaccine development. Acta Virol 61(3):377–390CrossRefPubMedPubMedCentralGoogle Scholar
  7. Golshani M, Buozari S (2017) A review of Brucellosis in Iran: epidemiology, risk factors, diagnosis, control, and prevention. Iranian Biomed J 21(6):349Google Scholar
  8. Jahandar MH, Forouharmehr A (2018) Optimization of human serum albumin periplasmic localization in Escherichia coli using in silico evaluation of different signal peptides. Int J Peptide Res Ther 1–9Google Scholar
  9. Jamshidi A, Razmyar J, Borji S (2014) Identification of Coxiella burnetii by touch-down PCR assay in unpasteurized milk and dairy products in North-East of Iran. Iranian J Veterin Med 8(1):15–19Google Scholar
  10. Kováčová E, Kazar J (2002) Q fever-still a query and underestimated infectious disease. Acta Virol 46:193–210PubMedGoogle Scholar
  11. Nazifi N, Tahmoorespur M, Sekhavati MH, Haghparast A (2018) Engineering, cloning and expression of DNA sequence coding of OMP31 epitope of Brucella melitensis linked to IL-2 in Escherichia coli. Int J Infect 5(3):e68974CrossRefGoogle Scholar
  12. Reeves PM, Paul SR, Sluder AE, Brauns TA, Poznansky MC (2017) Q-vaxcelerate: a distributed development approach for a new Coxiella burnetii vaccine. Hum Vac Immunother 13(12):2977–2981CrossRefGoogle Scholar
  13. Scheerlinck J-PY (2001) Genetic adjuvants for DNA vaccines. Vaccine 19(17–19):2647–2656CrossRefPubMedPubMedCentralGoogle Scholar
  14. Seshadri R, Paulsen IT, Eisen JA, Read TD, Nelson KE, Nelson WC et al (2003) Complete genome sequence of the Q-fever pathogen Coxiella burnetii. Proc Natl Acad Sci 100(9):5455–5460CrossRefPubMedPubMedCentralGoogle Scholar
  15. Singh M, O’Hagan D (1999) Advances in vaccine adjuvants. Nat Biotechnol 17(11):1075CrossRefPubMedPubMedCentralGoogle Scholar
  16. Vartak A, Sucheck SJ (2016) Recent advances in subunit vaccine carriers. Vaccines 4(2):12CrossRefGoogle Scholar
  17. Vigil A, Ortega R, Nakajima-Sasaki R, Pablo J, Molina DM, Chao CC et al (2010) Genome-wide profiling of humoral immune response to Coxiella burnetii infection by protein microarray. Proteomics 10(12):2259–2269CrossRefPubMedPubMedCentralGoogle Scholar
  18. Xiong X, Wang X, Wen B, Graves S, Stenos J (2012) Potential serodiagnostic markers for Q fever identified in Coxiella burnetii by immunoproteomic and protein microarray approaches. BMC Microbiol 12(1):35CrossRefPubMedPubMedCentralGoogle Scholar
  19. Yousefi S, Tahmoorespur M, Sekhavati MH (2016) Cloning, expression and molecular analysis of Iranian Brucella melitensis Omp25 gene for designing a subunit vaccine. Res Pharm Sci 11(5):412CrossRefPubMedPubMedCentralGoogle Scholar
  20. Zhang G, Samuel JE (2004) Vaccines against Coxiella infection. Expert Rev Vac 3(5):577–584CrossRefGoogle Scholar
  21. Zhang J, Yun J, Shang Z, Zhang X, Pan B (2009) Design and optimization of a linker for fusion protein construction. Prog Nat Sci 19(10):1197–1200CrossRefGoogle Scholar
  22. Zhang C, Wang B, Wang M. GM-CSF (2011) and IL-2 as adjuvant enhance the immune effect of protein vaccine against foot-and-mouth disease. Virol J 8(1):7CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Amin Jaydari
    • 1
  • Peyman Esmaeili Fard Barzegar
    • 1
  • Ali Forouharmehr
    • 2
  • Ali Kakanezhadifard
    • 1
  • Narges Nazifi
    • 3
  1. 1.Department of Microbiology, Faculty of Veterinary MedicineLorestan UniversityKhorramabadIran
  2. 2.Department of Animal Science, Faculty of AgricultureLorestan UniversityKhorramabadIran
  3. 3.Department of Animal Science, Faculty of AgricultureFerdowsi University of MashhadMashhadIran

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