Characterization of a Mycobacterium bovis BCG insertion sequence related to the IS21 family

Lefèvre, Philippe

doi:10.1016/s0378-1097(99)00359-6

Cited by 5 publications

(4 citation statements)

References 8 publications

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“…ORF1 overlaps 118 bp of the M. leprae 13B3 DNA fragment previously cloned by Mustafa et al (29) and 338 bp of the lppx gene of M. tuberculosis (30). The complete 833 bp ORF2, in the middle of the cloned fragment, encodes a protein which is 42% similar to the IstB subunit of E. coli IS21 (25). ORF3 at the other extremity of C3-1 had a size of 444 bp and was also incomplete.…”

Section: Screening Of a Gt11 M Bovis Bcg Genomic Library With Mabs Dmentioning

confidence: 87%

“…The fluorescence intensity was increased more than 10-fold compared to control staining with the secondary antibody only. It is not entirely clear whether the gene encoding the 22-kDa protein is present in other mycobacterial species, but indirect evidence based on PCR and insertion element analysis indicates that the gene can be found only in mycobacteria belonging to the M. tuberculosis complex and in M. leprae (25). Direct 22-kDa antigen detection based on flow cytometry, particle counting immunoassay (11), or dot blotting (3) using monoclonal and/or polyclonal antibodies against this strongly B-cell immunogenic protein could then be used for a rapid and cheap diagnosis of bacteria belonging to the M. tuberculosis complex.…”

Section: Discussionmentioning

confidence: 99%

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Cloning of the Gene Encoding a 22-Kilodalton Cell Surface Antigen ofMycobacterium bovisBCG and Analysis of Its Potential for DNA Vaccination against Tuberculosis

Lefèvre¹,

Denis²,

Wit³

et al. 2000

Infect Immun

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Using spleen cells from mice vaccinated with live Mycobacterium bovis BCG, we previously generated three monoclonal antibodies reactive against a 22-kDa protein present in mycobacterial culture filtrate (CF) (K. Huygen et al., Infect. Immun. 61:2687-2693, 1993). These monoclonal antibodies were used to screen an M. bovis BCG genomic library made in phage gt11. The gene encoding a 233-amino-acid (aa) protein, including a putative 26-aa signal sequence, was isolated, and sequence analysis indicated that the protein was 98% identical with the M. tuberculosis Lppx protein and that it contained a sequence 94% identical with the M. leprae 38-mer polypeptide 13B3 recognized by T cells from killed M. leprae-immunized subjects. Flow cytometry and cell fractionation demonstrated that the 22-kDa CF protein is also highly expressed in the bacterial cell wall and membrane compartment but not in the cytosol. C57BL/6, C3H, and BALB/c mice were vaccinated with plasmid DNA encoding the 22-kDa protein and analyzed for immune response and protection against intravenous M. tuberculosis challenge. Whereas DNA vaccination induced elevated antibody responses in C57BL/6 and particularly in C3H mice, Th1-type cytokine response, as measured by interleukin-2 and gamma interferon secretion, was only modest, and no protection against intravenous M. tuberculosis challenge was observed in any of the three mouse strains tested. Therefore, the 22-kDa antigen seems to have little potential for a DNA vaccine against tuberculosis, but it may be a good candidate for a mycobacterial antigen detection test.Tuberculosis remains a widespread and lethal infectious disease affecting millions of people worldwide. The World Health Organization estimates that there are about 8 million new cases each year and that the disease is responsible for at least 3 million deaths annually (5,12,23,34). The eradication of tuberculosis by early diagnosis, efficient therapy, and effective prophylactic vaccination remains an important goal. For the latter purpose, characterization of purified antigens implicated in protective immunity against mycobacterial infections is essential. The protective antigens of Mycobacterium tuberculosis are still not precisely defined, but several observations in experimental animal models have indicated that they predominantly reside within a pool of secreted or surface-exposed proteins that can be found in mycobacterial culture filtrate (CF) (2,15,16). Using the powerful technique of DNA vaccination, we and others have shown so far that at least four antigens present in M. tuberculosis CF are able to induce a protective immunity in mice against M. tuberculosis H37Rv infection: the 38-kDa PstS-1 and 40-kDa PstS-3 phosphate binding proteins (36, 41) and the 30-and 32-kDa components of the antigen 85 complex (17,22). Protective immune responses can also be induced with plasmid DNA encoding the 65-kDa heat shock protein, which is generally considered to be a cytosolic protein but is very abundant in CF from stressed mycobacterial cultures (37)...

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Section: Screening Of a Gt11 M Bovis Bcg Genomic Library With Mabs Dmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Cloning of the Gene Encoding a 22-Kilodalton Cell Surface Antigen ofMycobacterium bovisBCG and Analysis of Its Potential for DNA Vaccination against Tuberculosis

Lefèvre¹,

Denis²,

Wit³

et al. 2000

Infect Immun

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“…3b), and corresponded to a truncated Ist‐B like ATP‐binding protein described in the rapid grower mycobacteria Mycobacterium vanbaalenii PYR‐1 (Khan et al , 2002). Lower levels of similarity were found with the same Ist‐B‐like protein from other mycobacterial genomes, such as those of M. tuberculosis and Mycobacterium bovis BCG (Gordon et al , 1999; Lefevre et al , 1999). The Ist‐B protein of the M. tuberculosis H37Rv genome sequence is related to the transposase of the IS 1535 (Gordon et al , 1999), a member of the IS 607 family of ISs (http://www-is.biotoul.fr).…”

Section: Resultsmentioning

confidence: 99%

Molecular features ofMycobacterium aviumhuman isolates carrying a single copy of IS1245and IS1311per genome

Murcia

García

Otal

et al. 2007

FEMS Microbiology Letters

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Human clinical isolates of the Mycobacterium avium complex, from hospitals in Bogotá, were studied using a wide range of molecular tests including PCR restriction-enzyme analysis (PRA) of the hsp65 gene. Up to 21 of the isolates were identified as M. avium PRA variant III (Mav III), a variant obtained only from isolates on the American continent. In contrast to previous reports, restriction fragment length polymorphism analysis using IS1245 and IS1311 showed a single copy for each insertion sequence (IS) in the majority (19/21) of the Colombian Mav III isolates under study. In order to analyse whether the ISs were inserted in a relevant genomic region, experimental conditions were established to determine the insertion loci of each single copy of both ISs in the genome. Analysis of genomic insertion loci indicated that both IS1245 and IS1311 were present in areas containing putatively truncated integrases and/or transposases, which may have an influence on the mobility of the inserted IS. In addition, a conserved genomic region was identified for the insertion of IS1311; this region could be part of the IS1311 itself.

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“…xyli com a sonda R3/R4 após digestão eletrônica com as enzimas Bam HI, EcoRI, EcoRV e PstI. Os fragmentos marcados com "X" correspondem àqueles que também fo ram visualizados na hibridização do DNA genômico com a mesma sonda A sonda R5/R6 apresenta 52% de identidade a ISTB de Mycobacterium bovis(Lefèvre et al, 1999) e revelou 5 cópias desta seqüência quando hibridizadas ao DNA genômico clivado com as mesmas enzimas de restrição (Figura 12). O tamanho dos fragmentos gerados pela hibridização também coincide com aqueles obtidos nas análises eletrônicas com BamHI que também apresentou 5 cópias (Quadro 3).…”

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Análise estrutural e comparativa do genoma de Leifsonia xyli subsp. xyli

Gagliardi¹

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Characterization of a Mycobacterium bovis BCG insertion sequence related to the IS21 family

Cited by 5 publications

References 8 publications

Cloning of the Gene Encoding a 22-Kilodalton Cell Surface Antigen ofMycobacterium bovisBCG and Analysis of Its Potential for DNA Vaccination against Tuberculosis

Cloning of the Gene Encoding a 22-Kilodalton Cell Surface Antigen ofMycobacterium bovisBCG and Analysis of Its Potential for DNA Vaccination against Tuberculosis

Molecular features ofMycobacterium aviumhuman isolates carrying a single copy of IS1245and IS1311per genome

Análise estrutural e comparativa do genoma de Leifsonia xyli subsp. xyli

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