伯氏疏螺旋体外膜蛋白C基因在大肠杆菌中的表达

Expression of outer surface protein Cof Borrelia burgdorferi in Escherichia coli

目的 体外表达伯氏疏螺旋体外膜蛋白Ｃ（ＯｓｐＣ） ，以制备基因工程重组抗原用于莱姆病血清学诊断研究。方法 应用聚合酶链反应技术和基因重组技术，从２ 个伯氏疏螺旋体中国分离株基因组ＤＮＡ 中扩增得到ＯｓｐＣ基因，分别克隆到表达载体ＬＫＢ２ 中，在大肠杆菌中进行表达。结果 以天然蛋白形式高效表达了ＯｓｐＣ。扫描分析显示，表达的ＯｓｐＣ 相对分子质量均在２３ ０００ 左右，表达量占菌体总蛋白的３４ ％ ～５０ ％ 。Ｗｅｓｔｅｒｎ ｂｌｏｔ 试验证明，重组ＯｓｐＣ 蛋白可与莱姆病患者血清发生特异性反应。结论 重组ＯｓｐＣ

Objective In ordertoexpressoutersurface protein C(OspC) of Borrelia burgdorferiin vitro and make research on the serodiagnosis of Lime disease with recombinant protein . Methods OspC gene,amplified from two Chinese isolates of Borrelia burgdorferi, were cloned into an expression vector LKB2 respectively .Two expressed vectors BT OspCand BJ OspC were constructed and then expressedin E.coli.Results OspCproteins were expressed heavily in a nonfused form which weighted 23 000 and accounted for 34 % ～50 % oftotalbacterial protein . Western blot analysis showed thatthe expressed pro tein could bespecificallyrecognized bytheserafrom patients with Lyme disease.Conclusion Thesuccess fulexpression of recombinant OspCprotein makes a foundation for developing early diagnostic agents for Lyme disease.