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Cloning and expressing of the lipoprotein LppC from China strains Mycoplasma mycoide subsp. Mycoides SC
Author(s): 
Pages: 90-93
Year: Issue:  2
Journal: CHINESE JOURNAL OF PREVENTIVE VETERINARY MEDICINE

Keyword:  牛传染性胸膜肺炎(CBPP)丝状支原体丝状亚种SC型(MmmSC)LppC;
Abstract: 丝状支原体丝状亚种SC型是A类烈性传染病牛传染性胸膜肺炎的病原体.脂蛋白LppC是重要的免疫优势抗原,LppC基因是MmmSC的特异性基因.本研究通过PCR方法,扩增MmmSC标准株PG1的LppC N末端基因.将该序列克隆到pMDT-18载体上并测序,用pET32a为载体构建重组质粒进行诱导表达.LppC N末端基因长度为644 bp,将其与NCBI上发布的Afade株、欧洲群代表株L2比较,它的核酸序列同源性为100%,表明LppC基因在种内高度保守.原核表达结果得到大小为45 ku的目的蛋白,用Ni-NTA系统进行纯化得到高纯度的蛋白,Western blot结果表明,重组蛋白能与牛传染性胸膜肺炎阳性血清反应,具有免疫学活性,为进一步与LppQ融合表达奠定基础.
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