Wen J P, Eder C, Lipps H J
Institut für Zellbiologie, Universität Witten/Herdecke, Witten, Germany.
Nucleic Acids Res. 1995 May 25;23(10):1704-9. doi: 10.1093/nar/23.10.1704.
We describe the construction of a vector carrying the micronuclear versions of two macronuclear DNA molecules, one of which was modified by the insertion of a polylinker sequence. This vector was injected into the polytene chromosomes of the developing macronucleus of Stylonychia and its processing during further macronuclear development and its fate in the mature macronucleus were analyzed. In up to 30% of injected cells the modified macronuclear DNA sequence could be detected. While the internal eliminated sequences (IES) present in the macronuclear precursor DNA sequence are still retained in the mature macronucleus, the modified macronuclear DNA sequence is correctly cut out from the vector, telomeres are added de novo and it is stably retained in the macronucleus during vegetative growth of the cells. This vector system represents an experimental system that allows the identification of DNA sequences involved in the processing of macronuclear DNA sequences during macronuclear development.
我们描述了一种载体的构建,该载体携带两个大核DNA分子的微核版本,其中一个通过插入多克隆位点序列进行了修饰。将此载体注入嗜热栖热放线菌发育中的大核的多线染色体中,并分析其在进一步的大核发育过程中的加工过程及其在成熟大核中的命运。在高达30%的注射细胞中,可以检测到修饰的大核DNA序列。虽然大核前体DNA序列中存在的内部消除序列(IES)仍保留在成熟大核中,但修饰的大核DNA序列从载体中被正确切除,端粒被重新添加,并且在细胞营养生长期间它稳定地保留在大核中。该载体系统代表了一种实验系统,可用于鉴定在大核发育过程中参与大核DNA序列加工的DNA序列。