Dymshits G M, Rumiantseva G V, Frumgarts L A, Gruzdev A D, Zaĭniev G A
Mol Biol (Mosk). 1981 Jan-Feb;15(1):86-95.
The possibility of site-specific scission of DNA in polytene chromosomes in situ by means of the method of complementarily addressed fragmentation is demonstrated. The fragmentation of polytene chromosomes of Chironomus thummi resulting from the alkylation of the denatured chromosomes with oligoadenylate derivatives, followed by scission of the DNA in specific sites and enlargement of nicks with exonucleases was investigated. Single-stranded regions were registered by means of luminescence microscopy after staining the chromosomes with acridine orange. The pattern of degradation of the chromosomes depends on the length of the oligoadenylate part of the reagents and is different from that obtained with uridine-5'-methylphosphate alkylating derivative which does not form any complexes with DNA. Oligoadenylates inhibit the action of the alkylating derivatives on the chromosomes.
通过互补寻址断裂法在原位多线染色体中实现DNA位点特异性切割的可能性得到了证实。研究了经寡腺苷酸衍生物对变性染色体进行烷基化处理,随后在特定位点切割DNA并利用核酸外切酶扩大切口后,嗜菌摇蚊多线染色体的断裂情况。在用吖啶橙对染色体进行染色后,通过发光显微镜记录单链区域。染色体的降解模式取决于试剂中寡腺苷酸部分的长度,并且与用不与DNA形成任何复合物的尿苷-5'-甲基磷酸烷基化衍生物所获得的模式不同。寡腺苷酸可抑制烷基化衍生物对染色体的作用。