Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius
| Autor: | Lorraine Endersen, Helge Weingart, Christopher D Johnston, Daniel Pletzer, Rodney Govender, Aidan Coffey, John P. Bannantine, Roy D. Sleator, Jim O'Mahony |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Microbiology (medical)
Cytoplasm Recombinant Fusion Proteins Immunology lcsh:QR1-502 Reading frame Gene Expression Paratuberculosis Enzyme-Linked Immunosorbent Assay Microbiology lcsh:Microbiology MAP antigens codon optimization Epitopes Antigen Genes Reporter medicine Original Research Article Codon MAP vaccine Antigens Bacterial biology MMP Lactobacillus salivarius Antibodies Monoclonal Gene Expression Regulation Bacterial MptD biology.organism_classification medicine.disease Antibodies Bacterial Molecular biology Mycobacterium avium subsp. paratuberculosis Lactobacillus Protein Transport Open reading frame Infectious Diseases paratuberculosis Membrane protein expression host Codon usage bias Heterologous expression Johne's disease |
| Zdroj: | Frontiers in Cellular and Infection Microbiology, Vol 4 (2014) Frontiers in Cellular and Infection Microbiology |
| ISSN: | 2235-2988 |
| DOI: | 10.3389/fcimb.2014.00120 |
| Popis: | It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP) proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants were preserved in synthetic MMP proteins as shown by monoclonal antibody mediated ELISA. Moreover, MMP is a membrane protein in MAP, which is also targeted to the cellular surface of recombinant L. salivarius at levels comparable to MAP. Additionally, we previously engineered MAP3733c (encoding MptD) and show herein that MptD displays the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adheres to the MptD-specific bacteriophage fMptD. This work demonstrates there is potential for L. salivarius as a viable antigen delivery vehicle for MAP, which may provide an effective mucosal vaccine against Johne's disease. |
| Databáze: | OpenAIRE |
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