Codon optimisation to improve expression of a Mycobacterium avium ssp. paratuberculosis‐specific membrane‐associated antigen by Lactobacillus salivarius. Issue 1 (18th April 2013)

• Record Type: Journal Article
• Authors: Johnston, Christopher; Douarre, Pierre E.; Soulimane, Tewfik; Pletzer, Daniel; Weingart, Helge; MacSharry, John; Coffey, Aidan; Sleator, Roy D.; O'Mahony, Jim
• Is Part Of: Pathogens and disease. Volume 68:Issue 1(2013:Jun.)
• Page Start: 27
• Page End: 38
• Publication Date: 2013-04-18
• Subjects: Medical microbiology -- Periodicals; Pathogenic microorganisms -- Periodicals; Communicable diseases -- Microbiology -- Periodicals; Communicable diseases -- Pathogenesis -- Periodicals; Host-parasite relationships -- Periodicals; Systems biology -- Periodicals; 616.904105
• DOI: 10.1111/2049-632X.12040
• Languages: English
• ISSNs: 2049-632X

Abstract

<abstract abstract-type="main" id="fim12040-abs-0001"> <title>Abstract</title> <p>Subunit and DNA‐based vaccines against <italic>Mycobacterium avium</italic> ssp. <italic>paratuberculosis</italic> (MAP) attempt to overcome inherent issues associated with whole‐cell formulations. However, these vaccines can be hampered by poor expression of recombinant antigens from a number of disparate hosts. The high G+C content of MAP invariably leads to a codon bias throughout gene expression. To investigate if the codon bias affects recombinant MAP antigen expression, the open reading frame of a MAP‐specific antigen MptD (MAP3733c) was codon optimised for expression against a <italic>Lactobacillus salivarius</italic> host. Of the total 209 codons which constitute MAP3733c, 172 were modified resulting in a reduced G+C content from 61% for the native gene to 32.7% for the modified form. Both genes were placed under the transcriptional control of the PnisA promoter; allowing controlled heterologous expression in <italic>L. salivarius</italic>. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C‐termini of the two <italic>MptD</italic> genes. A > 37‐fold increase in expression was observed for the codon‐optimised MAP3733synth variant over the native gene. Due to the low cost and improved expression achieved, codon optimisation significantly improves the potential of <italic>L</italic>. <italic>salivarius</italic> as an oral vaccine stratagem against Johne's disease.</p> </abstract>