丝状支原体丝状亚种SC型LppC基因的表达纯化  

Cloning and expressing of the lipoprotein LppC from China strains Mycoplasma mycoide subsp. Mycoides SC

作　　者：李媛[1] 乔祖健[1] 王亮[1] 张建华[1] 辛九庆[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第2期90-93,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：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LppCN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。LppCN末端基因长度为644bp,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45ku的目的蛋白,用Ni-NTA系统进行纯化得到高纯度的蛋白,Westernblot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。Lipoprotein LppC is an immunodominant antigen of Mycoplasma mycoides subsp. Mycoides small-colony type （Mmm SC） which causes contagious bovine pleuropneumonia （CBPP）. In this study, the LppC gene was amplified from srtain of MmmSC PGI, cloned into pMDT-18 vector and sequenced. The LppC was 644 bp in length and shared 100 % sequence homology with those of Afade, L2 and PGl strains, indicating that LppC gene is highly conserved in MmmSC. The LppC fragment was then subcloned into expression vector pET32a for expression. A recombinant protein of 45 ku was obtained. The protein was purified using a Ni-NTA system and could react with CBPP positive bovine antiserum.

关 键 词：牛传染性胸膜肺炎(CBPP) 丝状支原体丝状亚种SC(MmmSC) LppC 

分 类 号：S852.62[农业科学—基础兽医学]