Lorenz M G, Wackernagel W
Fachbereich Biologie, Universität Oldenburg, Federal Republic of Germany.
Appl Environ Microbiol. 1987 Dec;53(12):2948-52. doi: 10.1128/aem.53.12.2948-2952.1987.
Adsorption and desorption of DNA and degradation of adsorbed DNA by DNase I were studied by using a flowthrough system of sand-filled glass columns. Maximum adsorption at 23 degrees C occurred within 2 h. The amounts of DNA which adsorbed to sand increased with the salt concentration (0.1 to 4 M NaCl and 1 mM to 0.2 M MgCl2), salt valency (Na+ less than Mg2+ and Ca2+), and pH (5 to 9). Maximum desorption of DNA from sand (43 to 59%) was achieved when columns were eluted with NaPO4 and NaCl for 6 h or with EDTA for 1 h. DNA did not desorb in the presence of detergents. It is concluded that adsorption proceeded by physical and chemical (Mg2+ bridging) interaction between the DNA and sand surfaces. Degradability by DNase I decreased upon adsorption of transforming DNA. When DNA adsorbed in the presence of 50 mM MgCl2, the degradation rate was higher than when it adsorbed in the presence of 20 mM MgCl2. The sensitivity to degradation of DNA adsorbed to sand at 50 mM MgCl2 decreased when the columns were eluted with 0.1 mM MgCl2 or 100 mM EDTA before application of DNase I. This indicates that at least two types of DNA-sand complexes with different accessibilities of adsorbed DNA to DNase I existed. The degradability of DNA adsorbed to minor mineral fractions (feldspar and heavy minerals) of the sand differed from that of quartz-adsorbed DNA.
采用填充沙子的玻璃柱流通系统研究了DNA的吸附与解吸以及吸附的DNA被DNase I降解的情况。在23℃下,最大吸附在2小时内发生。吸附到沙子上的DNA量随盐浓度(0.1至4M NaCl和1mM至0.2M MgCl2)、盐价(Na+ < Mg2+和Ca2+)以及pH(5至9)的增加而增加。当用NaPO4和NaCl洗脱柱子6小时或用EDTA洗脱1小时时,可实现DNA从沙子上的最大解吸(43%至59%)。在洗涤剂存在的情况下,DNA不会解吸。得出的结论是,吸附是通过DNA与沙子表面之间的物理和化学(Mg2+桥联)相互作用进行的。转化DNA吸附后,其被DNase I降解的能力下降。当DNA在50mM MgCl2存在下吸附时,降解速率高于在20mM MgCl2存在下吸附时的降解速率。在用DNase I处理之前,用0.1mM MgCl2或100mM EDTA洗脱柱子时,50mM MgCl2条件下吸附到沙子上的DNA对降解的敏感性降低。这表明至少存在两种类型的DNA - 沙子复合物,其吸附的DNA对DNase I的可及性不同。吸附到沙子的次要矿物组分(长石和重矿物)上的DNA的降解能力与吸附到石英上的DNA不同。