title: 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.
authors.affiliation: Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Namfon
authors.initials: N
authors.lastname: Suebwongsa
authors.affiliation: College of Medicine and Public Health, Ubon Ratchathani University, Ubon Ratchathani, Thailand.
authors.forename: Marutpong
authors.initials: M
authors.lastname: Panya
authors.affiliation: Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Wises
authors.initials: W
authors.lastname: Namwat
authors.affiliation: Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Saovaluk
authors.initials: S
authors.lastname: Sookprasert
authors.affiliation: Dairy Institute of Asturias, Villaviciosa, Spain.
authors.forename: Begoña
authors.initials: B
authors.lastname: Redruello
authors.affiliation: Dairy Institute of Asturias, Villaviciosa, Spain.
authors.forename: Baltasar
authors.initials: B
authors.lastname: Mayo
authors.affiliation: Dairy Institute of Asturias, Villaviciosa, Spain.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Alvarez
authors.affiliation: Department of Microbiology and Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
authors.forename: Viraphong
authors.initials: V
authors.lastname: Lulitanond
mesh_terms: D003001:Cloning, Molecular; D003062:Codon; D015870:Gene Expression; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics*; D007780:Lacticaseibacillus casei / Q000235:genetics* / Q000378:metabolism; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D015202:Protein Engineering
chemical_list: D003062:Codon; D019590:Nucleocapsid Proteins