Expression of surface-bound nonstructural 1 (NS1) protein of influenza virus A H5N1 on Lactobacillus casei strain C1

T. S. Tan, S. Syed Hassan, Wei Boon Yap

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

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 NS1 gene was first amplified and fused to the pSGANC332 expression plasmid. The NS1 protein expression was carried out by Lact. casei strain C1. PCR screening and DNA sequencing confirmed the presence of recombinant pSG-NS1-ANC332 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 NS1 was expressed as a 49-kDa protein in association with the anchoring peptide. The yield was 1·325 ± 0·065 μg mg−1 of bacterial cells. Lactobacillus casei expressing the NS1 on its cell wall was red-fluorescently stained, but the staining was not observed on Lact. casei carrying the empty pSGANC332. The results implied that Lact. casei strain C1 is a promising host for the expression of surface-bound NS1 protein using the pSGANC332 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.

Original languageEnglish
Pages (from-to)446-451
Number of pages6
JournalLetters in Applied Microbiology
Volume64
Issue number6
DOIs
Publication statusPublished - 1 Jun 2017

Fingerprint

Lactobacillus casei
Influenza A virus
Plasmids
Cell Wall
Proteins
Bacterial Proteins
Influenza Vaccines
Orthomyxoviridae
DNA Sequence Analysis
Fluorescent Antibody Technique
Staining and Labeling
Bacteria
Polymerase Chain Reaction
Peptides
Genes

Keywords

  • cloning
  • influenza viruses
  • Lactobacillus casei
  • NS1
  • protein expression
  • pSGANC332

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

Cite this

Expression of surface-bound nonstructural 1 (NS1) protein of influenza virus A H5N1 on Lactobacillus casei strain C1. / Tan, T. S.; Syed Hassan, S.; Yap, Wei Boon.

In: Letters in Applied Microbiology, Vol. 64, No. 6, 01.06.2017, p. 446-451.

Research output: Contribution to journalArticle

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