Zainiev G A, Gruzdev A D, Reznik N A
Tsitologiia. 1977 Jul;19(7):739-45.
Compactness of eukaryotic, particularly polytene chromosomes pose difficulties for investigation of their functional organization. For example, 0.1 --1.0 micrometer thick bands of polytene chromosomes contain some dozens microns pieces of DNA molecules. Therefore the useful resolving power of autoradigraphical methods is reduced by 10--100 times of its upper limit. To overcome the mentioned difficulty, a new method has been developed which permits to attain the upper limit of resolution. An isolated polytene chromosome from salivary gland nucleus of Chironomus thummi larva is stretched by microneedles to obtain a bundle of oriented DNP-fibers. A previously chosen small region of the chromosome (band or puff) is stretched simultaneously in a transverse direction by a stick frame made of another chromosome. Electron microscopy of the preparation reveals a meshwork of DNP fibers as presented on fig. 6.
真核生物染色体,特别是多线染色体的紧密性给研究其功能组织带来了困难。例如,多线染色体中0.1 - 1.0微米厚的带含有几十微米长的DNA分子片段。因此,放射自显影方法的有效分辨能力降低到其上限的10 - 100倍。为克服上述困难,已开发出一种新方法,该方法能够达到分辨率的上限。从嗜菌摇蚊幼虫唾液腺核中分离出的一条多线染色体,用微针拉伸以获得一束定向的DNP纤维。用由另一条染色体制成的棒框同时在横向拉伸染色体上预先选定的小区域(带或胀泡)。制备物的电子显微镜检查显示出如图6所示的DNP纤维网络。