Jun QIAO, Qingling MENG, Mengfan QIAO, Cheng CHEN, Xingxing ZHANG, Lulu TIAN, Haiting LU, Shasha GONG, Sufang YANG, Xuepeng CAI

Mycoplasma ovipneumoniae’nın Genomik İfade Kütüphanesinden İmmünojenik Genlerin Belirlenmesi ve Karakterizasyonu

Mycoplasma ovipneumoniae, koyunlarda solunum sistemi hastalığına neden olan önemli bir patojendir. Günümüzde, M. ovipneumoniae’nin immün ilişkili antijenleri hala bilinmemektedir ve bu durum M. ovipneumoniae’ya karşı yeni aşıların geliştirilmesini önemli ölçüde sınırlamaktadır. Çalışmada immün ilişkili antijen genlerinin tanımlanması ve karakterizasyonu amacıyla, M. ovipneumoniae’ye karşı pozitif klonlar pozitif serum tarafından belirlenip taranarak M. ovipneumoniae’nin genomik ekspresyon kütüphanesi oluşturuldu. Sekans analizi, bu 10 klonun, sırasıyla P97-benzeri protein, P102-benzeri protein, translasyon başlatma faktörü (IF-1), Metiyonin aminopeptidaz (MAP) ve P56 membran proteinini kodlayan 5 farklı gen içerdiğini gösterdi. Sırasıyla IF-1, MAP ve P97-benzeri proteini içeren üç protein, E. coli’de eksprese edildi ve immünojenisitelerini doğrulamak için kuzuların immünizasyonunda kullanıldı. Hayvan immünizasyon testi, yeni protein MAP’ın güçlü bir immünojenisite sergilediğini, P97 benzeri proteinin ve IF-1’in immünojenisitesinin nispeten zayıf olduğunu doğruladı. İmmünojenik protein MAP’ın tanımlanması, M. ovipneumoniae enfeksiyonuna karşı serolojik tanı yöntemi ve alt ünite aşı geliştirilmesi için değerli bir potansiyel antijen adayı elde edilmesini sağladı.

Identification and Characterization of Immunogenic Genes from Genomic Expression Library of Mycoplasma ovipneumoniae

Mycoplasma ovipneumoniae is an important pathogen causing respiratory disease in sheep. At present, the immune-associated antigens of M. ovipneumoniae are still unknown, which significantly limits the development of new vaccines for M. ovipneumoniae. In order to identify and characterize the immune-associated antigen genes, genomic expression library of M. ovipneumoniae was constructed and identified, from which positive clones were recognized and screened by positive serum against M. ovipneumoniae. Sequence analysis showed that these 10 clones contained 5 different genes encoding P97-like protein, P102-like protein, Translation initiation factor (IF-1), Methionine aminopeptidase (MAP) and P56 membrane protein, respectively. Three proteins including IF-1, MAP and P97-like protein were expressed in E. coli and used to immunize lambs to verify their immunogenicity, respectively. Animal immunization test confirmed that the novel protein MAP displayed a strong immunogenicity, while the immunogenicity of P97-like protein and IF-1 were relatively weak. The identification of immunogenic protein MAP provided a potentially valuable antigen candidate for the development of serological diagnostic method and subunit vaccine against M. ovipneumoniae infection.

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