Ivic A, MacDougall J, Russell R R, Penn C W
School of Biological Sciences, University of Birmingham, U.K.
FEMS Microbiol Lett. 1991 Mar 1;62(2-3):189-93. doi: 10.1016/0378-1097(91)90156-5.
Agarose gel electrophoresis of whole genomic DNA of the oral spirochaete Treponema denticola has revealed a plasmid-like fraction. Purification and restriction enzyme analysis has confirmed the presence of a 2.6-kb circular plasmid, which has been mapped for restriction sites and cloned into the Escherichia coli plasmid pUC18. Southern blot analysis of genomic T. denticola DNA, using the plasmid as a probe, has shown that the plasmid is present only as an extra-chromosomal element. No plasmid-coded recombinant gene product from a PstI insert in pUC18 has been detected in host cells of E. coli by SDS-PAGE or immunoblotting with polyclonal immune rabbit serum to T. denticola. The discovery of this plasmid may provide a useful tool in the application of new molecular approaches in spirochaetal biology.
对口腔螺旋体齿垢密螺旋体的全基因组DNA进行琼脂糖凝胶电泳，结果显示出一种类似质粒的成分。纯化和限制性酶切分析证实存在一个2.6kb的环状质粒，该质粒已绘制出限制性酶切位点图谱并克隆到大肠杆菌质粒pUC18中。用该质粒作为探针，对齿垢密螺旋体基因组DNA进行Southern印迹分析，结果表明该质粒仅作为一种染色体外元件存在。通过SDS-PAGE或用针对齿垢密螺旋体的多克隆免疫兔血清进行免疫印迹，在大肠杆菌宿主细胞中未检测到来自pUC18中PstI插入片段的质粒编码重组基因产物。该质粒的发现可能为在螺旋体生物学中应用新的分子方法提供一个有用的工具。
