Cloning and expression of a codon-optimized gene encoding the infl uenza A virus nucleocapsid protein in Lactobacillus casei

Autores/as

  • Namfon Suebwongsa Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases (RCEID), Faculty of Medicine, Khon Kaen University, Khon Kaen
  • Marutpong Panya College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani
  • Wises Namwat Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases (RCEID), Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
  • Saovaluk Sookprasert Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases (RCEID), Faculty of Medicine, Khon Kaen University, Khon Kaen
  • Begoña Redruello Dairy Institute of Asturias (IPLA-CSIC), Villaviciosa
  • Baltasar Mayo Dairy Institute of Asturias (IPLA-CSIC), Villaviciosa
  • Miguel A. Álvarez Dairy Institute of Asturias (IPLA-CSIC), Villaviciosa
  • Viraphong Lulitanond Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases Faculty of Medicine, Khon Kaen University

Palabras clave:

Lactobacillus casei, lactic acid bacteria, infl uenza A virus, viral nucleocapsid proteins, heterologous expression, codon usage

Resumen

Lactic acid bacteria (LAB) species are envisioned as promising vehicles for the mucosal delivery of therapeutic and prophylactic molecules, including the development of oral vaccines. In this study, we report on the expression of a synthetic nucleocapsid (NP) gene of infl uenza A virus in Lactobacillus casei. The NP gene was re-designed based on the tRNA pool and the codon usage preference of L. casei BL23. The codon-optimized NP gene was then cloned and expressed in L. casei RCEID02 under the control of a constitutive promoter, that of the lactate dehydrogenase (ldh) gene. The synthetic NP gene was further expressed in L. casei EM116 under the control of an inducible promoter, that of the structural gene of nisin (nisA) from Lactococcus lactis. Based on Western blot analysis, the specifi c protein band of NP, with a molecular mass of 56.0 kDa, was clearly detected in both expression systems. Thus, our study demonstrates the success of expressing a codon-optimized infl uenza A viral gene in L. casei. The suitability of the recombinant LAB strains for immunization purposes is currently under evaluation. [Int Microbiol 2013; 16(2):93-101]

Keywords: Lactobacillus casei; lactic acid bacteria; infl uenza A virus; viral nucleocapsid proteins; heterologous expression; codon usage

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Número

Vol. 16 Núm. 2 (2013)

Sección

Research Articles

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