Cloning and expression of P67 protein of Mycoplasma leachii

Contagious caprine pleuropneumonia (CCPP) is a serious disease of goats, occasionally sheep and wild ruminants, caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp). The disease is characterized by severe serofibrinous pleuropneumonia, very high morbidity ($100%), and mortality (80-100%). CCPP affects goats in more than 40 countries of the world thereby posing a serious threat to goat farming around the globe. The characteristic clinical signs of CCPP are severe respiratory distress associated with sero-mucoid nasal discharge, coughing, dyspnea, pyrexia, pleurodynia, and general malaise. In later stages, severe lobar fibrinous pleuropneumonia, profuse fluid accumulation in pleural cavity, severe congestion of lungs and adhesion formation is observed. Mycoplasmal antigen interactions with host immune system and its role in CCPP pathogenesis are not clearly understood. CCPP is not a zoonotic disease. Diagnosis has overcome cumbersome and lengthy conventional tests involving culture, isolation, and identification by advanced serological (LAT, cELISA) or gene-based amplification of DNA (PCR, RFLP, and hybridization) and sequencing. The latex agglutination test (LAT) is rapid, simple, and better test for field and real-time diagnosis applicable to whole blood or serum and is more sensitive than the CFT and easier than the cELISA. Moreover, the studies on antibiotic sensitivity and exploration of novel antibiotics (fluoroquinolones, macrolides) can help in better therapeutic management besides preventing menace of antibiotic resistance. Re-visiting conventional prophylactic measures focussing on developing novel strain-based or recombinant vaccines using specific antigens (capsular or cellular) should be the most important strategy for controlling the disease worldwide.

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“…Thankappan et al. (2017) cloned and expressed P67 protein of Mycoplasma leachii which may have prophylactic value. Tesgera et al.…”

Section: Vaccinationmentioning

confidence: 99%

“…2014; Thankappan et al. 2017; Thiaucourt et al. 2018), antibiotics and antibiotic resistance, which is already in rise in different mycoplasma (Prats-van der Ham et al.…”

Section: Introductionmentioning

confidence: 99%

Contagious caprine pleuropneumonia – a comprehensive review

Yatoo

Parray

Bashir

et al. 2019

Veterinary Quarterly

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“…Vector pRham-SUMO was used to clone and sequence the nagH gene of C. chauvoei [ 16 ]. In another study, P67 gene of Mycoplasma leachii was cloned in pRham N-His SUMO Kan vector and transformed into competent Escherichia cloni 10G cells [ 17 ]. In this study, the pET-32a ligA construct was transformed into E. coli DH5α, an expression host that lacks T7 RNA polymerase gene.…”

Section: Discussionmentioning

confidence: 99%

Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coli DH5α system

Soman¹,

Mini²,

Joseph³

et al. 2018

Vet World

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AimThis study aims at cloning, sequencing, and phylogenetic analysis of a partial CDS of ligA gene in pET-32a - Escherichia coli DH5α system, with the objective of identifying the conserved nature of the ligA gene in the genus Leptospira.Materials and MethodsA partial CDS (nucleotide 1873 to nucleotide 3363) of the ligA gene was amplified from genomic DNA of Leptospira interrogans serovar Canicola by polymerase chain reaction (PCR). The PCR-amplified DNA was cloned into pET-32a vector and transformed into competent E. coli DH5α bacterial cells. The partial ligA gene insert was sequenced and the nucleotide sequences obtained were aligned with the published ligA gene sequences of other Leptospira serovars, using nucleotide BLAST, NCBI. Phylogenetic analysis of the gene sequence was done by maximum likelihood method using Mega 6.06 software.ResultsThe PCR could amplify the 1491 nucleotide sequence spanning from nucleotide 1873 to nucleotide 3363 of the ligA gene and the partial ligA gene could be successfully cloned in E. coli DH5α cells. The nucleotide sequence when analyzed for homology with the reported gene sequences of other Leptospira serovars was found to have 100% homology to the 1910 bp to 3320 bp sequence of ligA gene of L. interrogans strain Kito serogroup Canicola. The predicted protein consisted of 470 aminoacids. Phylogenetic analysis revealed that the ligA gene was conserved in L.interrogans species.ConclusionThe partial ligA gene could be successfully cloned and sequenced from E. coli DH5α cells. The sequence showed 100% homology to the published ligA gene sequences. The phylogenetic analysis revealed the conserved nature of the ligA gene. Further studies on the expression and immunogenicity of the partial LigA protein need to be carried out to determine its competence as a subunit vaccine candidate.

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“…It needs to be developed from local isolates of mycoplasma. Novel vaccine development may be focused on recombinant vaccines, DNA vaccines, mycoplasma specific lipoprotein and polysaccharide utilization (Nicholas et al, 2009;Kumar et al, 2009;Kumar et al, 2014;Thankappan et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Mycoplasmosis in Small Ruminants in India: A Review

Yatoo¹,

Kashmir²,

Parray³

et al. 2018

JEBAS

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Mycoplasmosis, the diseases caused by mycoplasma, are one of the challenging and continuous threats to small ruminant farming. They cause heavy morbidity (upto 100%), mortality (10-100%) and huge economic loss. Common diseases caused by mycoplasma organisms are contagious caprine pleuropneumonia (mostly in goats), contagious agalactia (both sheep and goats), atypical pneumonia (usually sheep), besides arthritis, mastitis, seminal vesiculitis, ampullitis, epididymitis, orchitis, urethritis, conjunctivitis and meningitis either alone or as classical syndromes. The common mycoplasma includes Mycoplasma mycoides subspecies mycoides, M. mycoides subsp. capri, M. capricolum subsp. capripneumoniae, M. capricolum subsp. capricolum, M. agalactiae, M. bovis and M. conjunctivae etc. Most of the mycoplasmas produce a conglomerate of clinical signs hampering diagnosis. The other main challenge posed by mycoplasma is the difficulty in isolation due to lack of

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Cloning and expression of P67 protein of Mycoplasma leachii

Cited by 3 publications

References 30 publications

Contagious caprine pleuropneumonia – a comprehensive review

Contagious caprine pleuropneumonia – a comprehensive review

Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a - Escherichia coli DH5α system

Mycoplasmosis in Small Ruminants in India: A Review

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