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Expression of surface‐bound nonstructural 1 ( NS 1) protein of influenza virus A H5N1 on Lactobacillus casei strain C1
Author(s) -
Tan T.S.,
Syed Hassan S.,
Yap W.B.
Publication year - 2017
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12738
Subject(s) - lactobacillus casei , plasmid , biology , recombinant dna , microbiology and biotechnology , virus , influenza a virus , virology , bacteria , gene , biochemistry , genetics
The study aimed to construct a recombinant Lactobacillus casei expressing the nonstructural ( NS ) 1 protein of influenza A virus H5N1 on its cell wall. The NS 1 gene was first amplified and fused to the pSGANC 332 expression plasmid. The NS 1 protein expression was carried out by Lact. casei strain C1. PCR screening and DNA sequencing confirmed the presence of recombinant pSG ‐ NS 1‐ ANC 332 plasmid in Lact. casei . The plasmid was stably maintained (98·94 ± 1·65%) by the bacterium within the first 20 generations without selective pressure. The NS 1 was expressed as a 49‐ kD a protein in association with the anchoring peptide. The yield was 1·325 ± 0·065 μ g mg −1 of bacterial cells . Lactobacillus casei expressing the NS 1 on its cell wall was red‐fluorescently stained, but the staining was not observed on Lact. casei carrying the empty pSGANC 332. The results implied that Lact. casei strain C1 is a promising host for the expression of surface‐bound NS 1 protein using the pSGANC 332 expression plasmid. Significance and Impact of the Study The study has demonstrated, for the first time, the expression of nonstructural 1 (NS1) protein of influenza A virus H5N1 on the cell wall of Lactobacillus casei using the pSGANC332 expression plasmid. Display of NS1 protein on the bacterial cell wall was evident under an immunofluorescence microscopic observation. Lactobacillus casei carrying the NS1 protein could be developed into a universal oral influenza vaccine since the NS1 is highly conserved among influenza viruses.