Sargent M G, Bennett M F, Burdett I D
J Bacteriol. 1983 Jun;154(3):1389-96. doi: 10.1128/jb.154.3.1389-1396.1983.
When lysates of Bacillus subtilis were treated with restriction endonucleases EcoRI or HindIII, almost all of the DNA was released from the major plasma membrane fraction that was sedimentable at low speed. However, a very small part of the released DNA, when centrifuged at high speed, appeared to be bound to small membrane fragments. On agarose gels, this material, prepared with either enzyme, contained only a small number of restriction fragments, and the DNA in the sample hybridized with 11 to 12 EcoRI or HindIII fragments of chromosomal DNA. This DNA was used after nick-translation to screen Charon 4A clone banks for phages containing membrane-bound fragments. One of these was studied in detail. Only a part (about 5 kilobases) of the region present in this clone is important in binding the DNA to the membrane subparticle.
当用限制性内切酶EcoRI或HindIII处理枯草芽孢杆菌的裂解物时,几乎所有的DNA都从低速可沉淀的主要质膜部分释放出来。然而,当高速离心时,释放出的DNA中有非常小的一部分似乎与小的膜片段结合。在琼脂糖凝胶上,用这两种酶制备的这种物质只含有少量的限制性片段,并且样品中的DNA与染色体DNA的11至12个EcoRI或HindIII片段杂交。这种DNA在缺口平移后用于筛选Charon 4A克隆文库,以寻找含有膜结合片段的噬菌体。其中一个被详细研究。该克隆中存在的区域只有一部分(约5千碱基)在将DNA与膜亚颗粒结合方面很重要。