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

Abstract
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 influenza 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 specific 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 influenza A viral gene in L. casei. The suitability of the recombinant LAB strains for immunization purposes is currently under evaluation.
AuthorsNamfon Suebwongsa, Marutpong Panya, Wises Namwat, Saovaluk Sookprasert, Begoña Redruello, Baltasar Mayo, Miguel A Alvarez, Viraphong Lulitanond
JournalInternational microbiology : the official journal of the Spanish Society for Microbiology (Int Microbiol) Vol. 16 Issue 2 Pg. 93-101 (Jun 2013) ISSN: 1139-6709 [Print] Switzerland
PMID24400527 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Codon
  • Nucleocapsid Proteins
Topics
  • Cloning, Molecular
  • Codon
  • Gene Expression
  • Influenza A Virus, H3N2 Subtype (genetics)
  • Lacticaseibacillus casei (genetics, metabolism)
  • Nucleocapsid Proteins (genetics, metabolism)
  • Promoter Regions, Genetic
  • Protein Engineering

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