Walker E M, Arnett J K, Heath J D, Norris S J
Department of Pathology, University of Texas Medical School, Houston 77030.
Infect Immun. 1991 Jul;59(7):2476-9. doi: 10.1128/iai.59.7.2476-2479.1991.
The genome size and chromosome conformation of Treponema pallidum subsp. pallidum, Nichols strain, were determined by contour-clamped homogeneous electric field electrophoresis, a pulsed-field gel electrophoresis technique. Digestion of T. pallidum subsp. pallidum DNA with the restriction endonucleases NotI and SpeI produced 12 and 26 fragments, respectively. Summation of the physical lengths of the fragments produced by NotI and SpeI cleavage yielded average sizes of 900 and 913 kbp, respectively, for the genome of T. pallidum subsp. pallidum. Contour-clamped homogeneous electric field electrophoresis of T. pallidum subsp. pallidum DNA exposed to 4 krads of gamma irradiation resolved a single band of 800 to 1,000 kbp; treatment of the DNA with 16 krads of gamma irradiation resulted in the production of smaller fragments, whereas untreated DNA did not migrate into the gels. The gamma irradiation results indicate that T. pallidum subsp. pallidum has a single, circular chromosome that was linearized at a dosage of 4 krads of gamma irradiation. The size estimate provided by restriction endonuclease digestion with NotI and SpeI shows that the genome of T. pallidum subsp. pallidum, at approximately 900 kbp, is considerably smaller than the 13,700-kbp genome size calculated from renaturation kinetics.
采用轮廓夹恒定电场电泳这一脉冲场凝胶电泳技术,测定了梅毒螺旋体苍白亚种Nichols株的基因组大小和染色体构象。用限制性内切酶NotI和SpeI消化梅毒螺旋体苍白亚种DNA,分别产生了12个和26个片段。NotI和SpeI切割产生的片段物理长度总和显示,梅毒螺旋体苍白亚种基因组的平均大小分别为900 kbp和913 kbp。对暴露于4 kradγ射线的梅毒螺旋体苍白亚种DNA进行轮廓夹恒定电场电泳,分离出一条800至1000 kbp的单带;用16 kradγ射线处理该DNA导致产生较小片段,而未处理的DNA未迁移到凝胶中。γ射线照射结果表明,梅毒螺旋体苍白亚种有一条单一的环状染色体,在4 kradγ射线剂量下被线性化。用NotI和SpeI进行限制性内切酶消化提供的大小估计表明,梅毒螺旋体苍白亚种的基因组大小约为900 kbp,比根据复性动力学计算的13700 kbp基因组大小小得多。