Codon optimisation to improve expression of a Mycobacterium avium ssp paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius

Typeset version

 

TY  - JOUR
  - Johnston, C,Douarre, PE,Soulimane, T,Pletzer, D,Weingart, H,MacSharry, J,Coffey, A,Sleator, RD,O'Mahony, J
  - 2013
  - June
  - Pathogens And Disease
  - Codon optimisation to improve expression of a Mycobacterium avium ssp paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius
  - Validated
  - ()
  - codon optimisation Mycobacterium avium Lactobacillus salivarius CONTROLLED GENE-EXPRESSION HETEROLOGOUS PROTEIN EXPRESSION COMPLETE GENOME SEQUENCE LACTIC-ACID BACTERIA SUBSP-PARATUBERCULOSIS ESCHERICHIA-COLI VACCINE DEVELOPMENT LACTOCOCCUS-LACTIS SYSTEM STRAIN
  - 68
  - 27
  - 38
  - Subunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (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 Lactobacillus salivarius 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 L.salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD 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 L.salivarius as an oral vaccine stratagem against Johne's disease.
  - 10.1111/2049-632X.12040
DA  - 2013/06
ER  - 
@article{V243944115,
   = {Johnston,  C and Douarre,  PE and Soulimane,  T and Pletzer,  D and Weingart,  H and MacSharry,  J and Coffey,  A and Sleator,  RD and O'Mahony,  J },
   = {2013},
   = {June},
   = {Pathogens And Disease},
   = {Codon optimisation to improve expression of a Mycobacterium avium ssp paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius},
   = {Validated},
   = {()},
   = {codon optimisation Mycobacterium avium Lactobacillus salivarius CONTROLLED GENE-EXPRESSION HETEROLOGOUS PROTEIN EXPRESSION COMPLETE GENOME SEQUENCE LACTIC-ACID BACTERIA SUBSP-PARATUBERCULOSIS ESCHERICHIA-COLI VACCINE DEVELOPMENT LACTOCOCCUS-LACTIS SYSTEM STRAIN},
   = {68},
  pages = {27--38},
   = {{Subunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (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 Lactobacillus salivarius 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 L.salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD 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 L.salivarius as an oral vaccine stratagem against Johne's disease.}},
   = {10.1111/2049-632X.12040},
  source = {IRIS}
}
AUTHORSJohnston, C,Douarre, PE,Soulimane, T,Pletzer, D,Weingart, H,MacSharry, J,Coffey, A,Sleator, RD,O'Mahony, J
YEAR2013
MONTHJune
JOURNAL_CODEPathogens And Disease
TITLECodon optimisation to improve expression of a Mycobacterium avium ssp paratuberculosis-specific membrane-associated antigen by Lactobacillus salivarius
STATUSValidated
TIMES_CITED()
SEARCH_KEYWORDcodon optimisation Mycobacterium avium Lactobacillus salivarius CONTROLLED GENE-EXPRESSION HETEROLOGOUS PROTEIN EXPRESSION COMPLETE GENOME SEQUENCE LACTIC-ACID BACTERIA SUBSP-PARATUBERCULOSIS ESCHERICHIA-COLI VACCINE DEVELOPMENT LACTOCOCCUS-LACTIS SYSTEM STRAIN
VOLUME68
ISSUE
START_PAGE27
END_PAGE38
ABSTRACTSubunit and DNA-based vaccines against Mycobacterium avium ssp. paratuberculosis (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 Lactobacillus salivarius 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 L.salivarius. Expression was monitored using fluorescence microscopy and microplate fluorometry via GFP tags translationally fused to the C-termini of the two MptD 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 L.salivarius as an oral vaccine stratagem against Johne's disease.
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URL
DOI_LINK10.1111/2049-632X.12040
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