Nagai K, Hendrickson W, Balakrishnan R, Yamaki H, Boyd D, Schaechter M
Proc Natl Acad Sci U S A. 1980 Jan;77(1):262-6. doi: 10.1073/pnas.77.1.262.
A complex consisting of replicative origin DNA and several proteins was isolated from Escherichia coli. Cells of temperature-sensitive mutants were labeled at the origin and fractionated by sucrose gradient centrifugation. A complex highly purified in origin DNA sedimented as a unique band. This complex dissociated at high concentration, above 0.2 M KCl. Upon dialysis, the complex reformed, allowing further purification of its constituents. Three major protein bands were found, corresponding to proteins of the outer membrane. The complex did not sediment with membrane fractions, but adhered to the outer membrane in the presence of magnesium.
从大肠杆菌中分离出一种由复制起点DNA和几种蛋白质组成的复合物。对温度敏感突变体的细胞在起点处进行标记,然后通过蔗糖梯度离心进行分级分离。在起点DNA中高度纯化的复合物沉降为一条独特的条带。该复合物在高于0.2 M KCl的高浓度下会解离。透析后,复合物会重新形成,从而可以进一步纯化其成分。发现了三条主要的蛋白条带,对应于外膜蛋白。该复合物不会与膜组分一起沉降,但在镁存在的情况下会附着在外膜上。