Ohnuki T, Imanaka T, Aiba S
Gene. 1983 Nov;25(1):155-9. doi: 10.1016/0378-1119(83)90178-6.
Thirty independent actinomycetes strains carrying plasmids were isolated from soil. These plasmids were purified as cccDNA by CsCl-EtBr equilibrium density-gradient centrifugation. Plasmids that induce "pocks", namely formation of circular zones of sporulation-inhibition, were selected by protoplast transformation of streptomycin-producing strain, Streptomyces griseus ATCC10137. Six pock-forming plasmids, pOA7, pOA11, pOA15, pOA23, pOA29 and pOA30, were obtained, and their cleavage maps, transformation frequencies, and copy numbers, as well as their stability, are described.
从土壤中分离出30株携带质粒的独立放线菌菌株。这些质粒通过氯化铯-溴化乙锭平衡密度梯度离心法纯化,得到共价闭合环状DNA(cccDNA)。通过对产链霉素菌株灰色链霉菌ATCC10137进行原生质体转化,筛选出能诱导“菌斑”(即形成孢子形成抑制的圆形区域)的质粒。获得了6种形成菌斑的质粒,即pOA7、pOA11、pOA15、pOA23、pOA29和pOA30,并描述了它们的酶切图谱、转化频率、拷贝数及其稳定性。