Genetic and antigenic characterization of the surface lipoprotein P48 of Mycoplasma bovis
Genetic and antigenic characterization of the surface lipoprotein P48 of Mycoplasma bovis
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DOI：
10.1016/j.vetmic.2005.05.007
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发表时间：
2005-08-30
影响因子：
3.3
通讯作者：
Rosati, S
中科院分区：
文献类型：
--
作者：
Robino, P;Alberti, A;Rosati, S
研究了无乳支原体P48同源膜脂蛋白在不同地理位置和生物学特性选择的牛支原体分离株中的存在。还讨论了其作为诊断工具的潜力。用兔抗血清检测无乳杆菌重组蛋白P48，在所有牛分枝杆菌分离株中观察到一个特异性信号，表明该蛋白在牛分枝杆菌簇中具有结构和抗原性保守性。在检测牛中发现的六种不同支原体时未检测到信号。采用PCR法对p48基因进行了鉴定和部分测序。采用细菌染色体直接测序法获得了基因的全长序列。将5个uga选择性突变为UGG，克隆出缺失信号肽的全长突变基因，并在大肠杆菌中表达。纯化后的重组抗原（r-P48）作为一种潜在的感染标记物进行了评估，该标记物使用了86种具有良好特征的实验和自然感染牛血清。实验感染后6-9天内检测到特异性IgM抗体，接触后第3 / 4周出现IgG应答。虽然抗体滴度远低于感染无乳分枝杆菌的绵羊或山羊，但结果表明牛分枝杆菌r-P48可以作为感染的特异性标记物。(C) 2005 Elsevier B.V.版权所有
The presence of a membrane lipoprotein homologous to the P48 of Mycoplasma agalactiae was investigated in different Mycoplasma bovis isolates selected by geographical locations and biological properties. Its potential as a diagnostic tool was also discussed. The presence of a specific signal observed in all M. bovis field isolates probed with a rabbit antiserum raised against the M. agalactiae recombinant P48 demonstrated that this protein is structurally and antigenically conserved within the M. bovis cluster. No signal was detected when testing six different mycoplasma species found in cattle. The p48 gene was identified by PCR approach and partially sequenced. Full length gene sequence was obtained by direct bacterial chromosome sequencing. Five UGAs were selectively mutated into UGG and the full length mutated gene, lacking the signal peptide, was cloned and expressed in Escherichia coli. The purified recombinant antigen (r-P48) was evaluated as a potential marker of infection using a panel of 86 well-characterized sera from experimentally and naturally infected cattle. Specific IgM antibodies were detected within 6-9 days after experimental infection followed by an IgG response lasting from the third/fourth week after contact.Although antibody titers were well below those observed in sheep or goats infected with M. agalactiae, results suggest that M. bovis r-P48 can be used as a specific marker of infection. (C) 2005 Elsevier B.V. All rights reserved.