Eiffert H, Schlott T, Hoppert M, Lotter H, Thomssen R
Department of Medical Microbiology, University of Göttingen, Germany.
J Med Microbiol. 1992 Mar;36(3):209-14. doi: 10.1099/00222615-36-3-209.
DNA of Borrelia burgdorferi was cleaved by the endonuclease EcoRI and ligated with the bacteriophage expression vector lambda gt11. After infection of the Escherichia coli strain Y1089, the plaques of recombinant phages were screened with a B. burgdorferi antiserum (human) for fusion proteins containing borrelia antigen.s A positive clone produced a hybrid protein (p200) of c. 200 Kda. The corresponding native borrelia protein (p97) was identified as having an Mr of 97 Kda. To localise protein p97 in the B. burgdorferi cell, immunoelectronmicroscopy and a Western blot of isolated flagella were used. Antibodies directed against proteins p200 and p97 recognised epitopes associated with the flagella.
伯氏疏螺旋体的DNA被核酸内切酶EcoRI切割，并与噬菌体表达载体λgt11连接。在用重组噬菌体感染大肠杆菌Y1089菌株后，用伯氏疏螺旋体抗血清（人）筛选含有疏螺旋体抗原的融合蛋白的噬菌斑。一个阳性克隆产生了一种约200 kDa的杂交蛋白（p200）。相应的天然疏螺旋体蛋白（p97）被鉴定为分子量为97 kDa。为了在伯氏疏螺旋体细胞中定位蛋白p97，使用了免疫电子显微镜和分离鞭毛的蛋白质印迹法。针对蛋白p200和p97的抗体识别与鞭毛相关的表位。
