摘要
目的原核表达伯氏疏螺旋体鞭毛蛋白Flagellin A基因特异性区段,获得重组鞭毛蛋白平截性蛋白作为诊断抗原,建立间接ELISA方法用于动物莱姆病的诊断。方法 PCR扩增获取伯氏疏螺旋体鞭毛蛋白基因的同源性较低的第394-798bp区段,构建重组质粒pGEX-4T-1/tFlaA,构建好的表达质粒转化到大肠杆菌BL21(DE3)中进行表达,并纯化重组蛋白,用纯化的表达蛋白作为莱姆病诊断的抗原,用于ELISA检测实验感染小鼠莱姆病。结果成功构建莱姆病螺旋体鞭毛平截性蛋白的表达载体,重组蛋白在宿主菌内高效、稳定表达,重组平截性蛋白显示了可作为ELISA诊断的抗原用于莱姆病的诊断价值。结论纯化的伯氏疏螺旋体鞭毛蛋白可作为莱姆病ELISA诊断抗原用于莱姆病的诊断,为莱姆病快速诊断试剂盒的开发打下基础。
The purpose was to extract Borrelia burgdorferi DNA and use PCR method to obtain the gene coding of the Borrelia burgdorferi flagellin from 394 to 798 nucleotide that exhibited low homology with related gene from other bacterial specises we insert Borrelia burgdorferi flagellin from 394 to 798 nucleotide to PGEX-4T-1 vector,verified by DNA sequence detection and tren thansformed it into in E.coli BL 21（DE3） to induce target protein.Then we purified the recombination protein as ELISA antigen to diagnosis Lyme disease.On other hand,we infected mouse with Borrelia burgdorferi.Two weeks,we got positive serum from mouse.We used recombinant protein as diagnostic antigen andpositive mouse serum as antibody to conduct ELISA test and analysis.The ELISA results showed the feasibility of recombinant protein as diagnostic antigen to diagnose Lyme disease,which lay foundations for the diagnosis of Lyme disease.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2011年第12期1106-1110,共5页
Chinese Journal of Zoonoses
