伯氏疏螺旋体鞭毛蛋白特异性区段的基因克隆和表达

GENE CLONE AND EXPRESSION OF THE SPECIFIC FRAGMENT OF FLAGELLIN FROM BORRELIA BURGDORFERI

目的表达伯氏疏螺旋体鞭毛蛋白特异性区段，探讨其作为诊断抗原的可行性。方法用ＰＣＲ方法获取与其它种属螺旋体编码鞭毛蛋白的基因序列同源性较低的第４０９－７８６ｂｐ区段，经ＤＮＡ序列测定证实后，经克隆、转化，在大肠杆菌ＢＬ２１（ＤＥ３）中表达，并做Ｗｅｓｔｅｒｎ－ｂｏｌｔ分析。结果重组蛋白在宿主菌内表达高效、稳定，Ｗｅｓｔｅｒｎ－ｂｌｏｔ示与抗鞭毛蛋白的单克隆抗体有较好的免疫反应性。结论以重组鞭毛蛋白的特异性区段作为诊断抗原具有可行性。

Aim Expressing the specific fragment of flagellin from Borrelia burgdorferi ,in order to evaluate the feasibility of this recombinant protein as a diagnostic antigen. Methods By using PCR method,to obtain the gene coding the Borrelia burgdorferi flagellin from 409bp to 786bp that exhibit low homology with related gene from other bacterial specise.Verified by DNA sequence detection,cloning and thansforming it was expressed in E.coli BL21(DE3)and evaluated by westernblot. Results The expressing of recombinant protein in host bacteria is effective and stable.Westernblot assay verified the immunological response to monoclone antidody to flagellin. Conclusion It showed the feasibility of recombinant protein as a diagnostic antigen.