Identification and characterization of a gene encoding a 35-kDa protein from<i>Mycobacterium avium</i>subspecies<i>paratuberculosis</i>

Banasure, K; Basagoudanavar, Suresh H.; Chaudhury, P.; Tiwari, Vivek; Parihar, N. S.; Goswami, P. P.

doi:10.1111/j.1574-6968.2001.tb10564.x

Cited by 14 publications

(4 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is notable that although MMP is not specific to MAP, with homologs existing in other mycobacterial species including M. avium (MAC) and M. leprae , previous genetic and serological evidence suggests that the protein is absent from M. tuberculosis and M. bovis (Triccas et al, 1998; Banasure et al, 2001). In addition, a DNA vaccine incorporating the M. leprae MMP-1 protein in isolation demonstrated significant levels of protective efficacy against M. leprae footpad infection in outbred Swiss Albino mice (Martin et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

Johnston

Bannantine

Govender

et al. 2014

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

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.

show abstract

Section: Introductionmentioning

confidence: 99%

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

Johnston

Bannantine

Govender

et al. 2014

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

show abstract

“…The mpt operon codes for mptA, mptB, mptC, mptD, mptE and mptF genes and among these, mptD codes for proteins exposed on the surface of MAP (Heinzmann et al 2008). Similarly, 35-kDa antigen gene codes for a strongly immunoreactive major membrane protein (MMP) and is associated with its virulence (Banasure et al 2001, Bannantine et al 2003. Since these genes could be candidates for developing newer diagnostics and vaccines, we generated recombinant mptD and 35 kDa antigens.…”

mentioning

confidence: 99%

“…These recombinant proteins can be of use in DIVA (Differentiating Infected from Vaccinated Animals) strategies. In this context, we cloned and expressed two important proteins of MAP namely, mptD and 35 kDa antigen which have been reported to be associated with virulence and hence could be of diagnostic and or immunogenic importance (Banasure et al 2001, Bannantine et al 2003, Stratmann et al 2004). The A260/A280 ratio of isolated MAP genomic DNA was found to be between 1.7 and 1.8 indicating the purity of the DNA as expected.…”

mentioning

confidence: 99%

“…Mpt operon clones of M. bovis BCG were found to reduce the amplification of MAP significantly in mice. Similarly, 35 kDa antigen gene codes for a strongly immunoreactive MMP that plays a role in invasion of epithelial cells and associated with the virulence of the organism (Banasure et al 2001, Bannantine et al 2003. Earlier work on the 35 kDa antigen protein had indicated that p35 recombinant protein has the potential for use in the serodiagnosis of animals with JD (El-Zaatari et al1996).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Expression of immunogenic genes of Mycobacterium avium subsp. paratuberculosis

Kumar

Vijayarani

Kumanan

2022

Indian J of Anim Sci

View full text Add to dashboard Cite

SUMMARY Johneâ€™s disease is an economically important disease caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Continuous efforts are being made to identify MAP genes with antigenic and immunogenic capabilities which may serve as vaccine candidates and diagnostic antigens. mptD and 35 KDa antigen genes of MAP amplified by Polymerase chain reaction resulted in 630bp and 830 bp products respectively. Both mptD and 35 KDa antigen genes were cloned in to pTZ57R/T cloning vector, and subsequently subcloned into pET22b and expressed in BL21 (DE3) host system. The expressed recombinant antigens could be tested in diagnostic assays and might open up the avenue for the development of newer diagnostics and vaccine candidates for Johneâ€™s disease.

show abstract

Identification of proteins of potential diagnostic value for bovine paratuberculosis

2006

View full text Add to dashboard Cite

Previously we showed that Mycobacterium paratuberculosis culture filtrates (CFs) contain more antigens that react with sera from infected cattle than do cellular extracts of the organism. The goal of the present study was to identify proteins of potential diagnostic value among these CF proteins. Proteins of potential interest were first separated by 2-DE. Roughly 240 CF protein spots were detected on CBB-stained gels using Phoretix 2D software. Of these, 83% reacted with serum from M. paratuberculosis-infected cattle in immunoblots. When bovine serum was absorbed with M. phlei antigens, however, only 37 of these antigenic protein spots were reactive. Twenty-four of these spots were selected for identification based on their immunoblot staining intensity and differences in pI and mass. A total of 14 proteins were ultimately identified by MS and BLAST searches as ModD, PepA, ArgJ, CobT, Antigen 85C, and nine hypothetical proteins. N-terminal peptide analysis of PepA, Antigen 85C, ModD, MAP1693c, MAP2168c, and MAP1022c showed that each protein has 27-39 amino acids that may function as a signal sequence suggesting they are secreted through a Sec-dependent pathway. These 14 proteins from M. paratuberculosis CF are strong candidates for use as antigens for improved serodiagnostic tests for bovine paratuberculosis.

show abstract

Identification and characterization of a gene encoding a 35-kDa protein fromMycobacterium aviumsubspeciesparatuberculosis

Cited by 14 publications

References 14 publications

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

Expression of immunogenic genes of Mycobacterium avium subsp. paratuberculosis

Identification of proteins of potential diagnostic value for bovine paratuberculosis

Contact Info

Product

Resources

About