Expression and antigenic characterization of recombinant Mycoplasma agalactiae P48 major surface protein

Rosati, Sergio; Robino, Patrizia; Fadda, Maria Elisabetta; Pozzi, Sarah; Mannelli, Alessandro; Pittau, Marco

doi:10.1016/s0378-1135(99)00164-9

Cited by 36 publications

(52 citation statements)

References 16 publications

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“…39,40 The presence of a 45-kD homologue (P45) of this protein in M. bovis has recently been demonstrated. 38 The use of a rabbit hyperimmune serum raised against this M. agalactiae antigen (␣-rP48) allowed us to establish its presence in all the M. bovis field strains studied and to confirm the identity of all 39 Mycoplasma colonies isolated during this study (collectively ascribed to strain Sar1) as M. bovis.…”

Section: Discussionmentioning

confidence: 99%

Molecular and Antigenic Characterization of aMycoplasma BovisStrain Causing an Outbreak of Infectious Keratoconjunctivitis

et al. 2006

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Abstract. An unusually high incidence of infectious keratoconjunctivitis followed by pneumonia and arthritis was observed in beef calves of a managed herd. No Moraxella spp. or bacteria other than Mycoplasma spp. were obtained from conjunctival and nasal swabs. A strategy was designed for characterization of bovine mycoplasmas at species and strain level on the basis of a combination of molecular tools and the immunoblotting method. The strategy made it possible to rapidly assign the bacterium responsible for this outbreak to one of the phylogenetic clusters of bovine mycoplasmas delineated in this study and then to identify it as Mycoplasma bovis. The strain, designated Sar 1, showed a 100% 16S rDNA sequence identity with two European strains (120/81 and MC3386) isolated in Germany and Ireland, respectively, and hosts a vsp gene analog to the vsp A , vsp , and vsp 422-8 genes of the M. bovis reference strain PG45 T and of the field strain 422. The use of a crossreactive rabbit serum developed against the Mycoplasma agalactiae immunodominant antigen P48 confirmed the molecular findings. The immunological response of calves against M. bovis was also investigated. This is the first report on the occurrence of M. bovis on the Island of Sardinia (Italy).

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Section: Discussionmentioning

confidence: 99%

Molecular and Antigenic Characterization of aMycoplasma BovisStrain Causing an Outbreak of Infectious Keratoconjunctivitis

et al. 2006

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“…The mammalian expression vectors pcDNA3.1 (Invitrogen), pVAX1 (Invitrogen), pCMV-Script (Stratagene), and the bacterial expression vector pGEX-2T/P48 (Rosati et al, 2000), were chosen to clone and express the P48 antigen gene of M. agalactiae. Briefly, the 1.3 kb DNA fragment containing the P48 gene, lacking the leader peptide, was obtained by PCR (Mastercycler Gradient, Eppendorf) from plasmid pGEX-2T/P48, using the primer pair MagP48/BamHI/K/ATG/F (5 0 -CTGGGATCCACGATGGTAAAAACTA TTTCAACACTTGCA-3 0 ) and MagP48/ECO/R (5 0 -CACGAA TTCTTATTTTCTTGTTTCAGAAGCCAA-3 0 ).…”

Section: Construction Of Expression Plasmidsmentioning

confidence: 99%

“…Escherichia coli strain JM105 was transformed with plasmid pGEX-2T/P48, and recombinant P48 was expressed and purified, as previously described (Rosati et al, 2000). Briefly, early log phase cultures of positive clones, induced for 2 h with 0.5 mM isopropyl b-thiogalactopyranoside (IPTG) were centrifuged and lysed by conventional physico-chemical methods.…”

Section: Expression and Purification Of P48 Antigenmentioning

confidence: 99%

“…We recently described the P48 of M. agalactiae as an invariable, constantly expressed, immunodominant, surface lipoprotein (Rosati et al, 1999) homologue to the MALP-404 lipoprotein of Mycoplasma fermentans (Davis and Wise, 2002). We also developed a commercial recombinant indirect ELISA for contagious agalactia (Institute Pourquier, France) based on the use of the P48 antigen lacking the leader peptide, which has been proven to be associated to high levels of specificity and sensitivity (Rosati et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

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Genetic immunization with the immunodominant antigen P48 of Mycoplasma agalactiae stimulates a mixed adaptive immune response in BALBc mice

Chessa

Pittau

Puricelli

et al. 2009

Research in Veterinary Science

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a b s t r a c tA DNA vaccine against contagious agalactia was developed for the first time, encoding the P48 of Mycoplasma agalactiae. Specific immune responses elicited in BALB/c mice were evaluated. Both total IgG and IgG1 were detected in mice vaccinated with pVAX1/P48. Proliferation of mononuclear cells of the spleen, levels of gamma interferon, interleukin-12, and interleukin-2 mRNAs were enhanced in immunized animals. Results indicate that pVAX1/P48 vaccination induced both T h 1 and T h 2 immune responses. Nucleic acid immunization could be a new strategy against M. agalactiae infections and may be potentially used to develop vaccines for other Mycoplasma diseases.

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“…Assim sendo, nestes organismos, a tradução de um mRNA de um gene de micoplasma vai ser interrompida em locais onde deveria ser incorporado um triptofano, resultando em proteínas truncadas (RAZIN et al, 1998). A produção de mutações sítio específicas no sentido de alterar os códons TGA por TGG, um códon padrão para triptofano em outros organismos, tem sido a estratégia utilizada para contornar este problema (CHUNG et al, 2000;ROSATI et al, 2000;NOH et al, 2002).…”

Section: Imunoprofilaxiaunclassified

Etiopatogenia e imunoprofilaxia da pneumonia enzoótica suína

Conceição

Dellagostin

2006

Cienc. Rural

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Expression and antigenic characterization of recombinant Mycoplasma agalactiae P48 major surface protein

Cited by 36 publications

References 16 publications

Molecular and Antigenic Characterization of aMycoplasma BovisStrain Causing an Outbreak of Infectious Keratoconjunctivitis

Molecular and Antigenic Characterization of aMycoplasma BovisStrain Causing an Outbreak of Infectious Keratoconjunctivitis

Genetic immunization with the immunodominant antigen P48 of Mycoplasma agalactiae stimulates a mixed adaptive immune response in BALBc mice

Etiopatogenia e imunoprofilaxia da pneumonia enzoótica suína

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