应用大肠杆菌染色素DNA在非洲爪蟾卵提取物实现诱导非细胞体系…

近年来我们实验室已成功地利用细胞核体外组装的实验模式，将多种生物的ＤＮＡ在非洲爪蟾卵提取物中实现了非细胞体系核装配。但亲缘关系最远的原核生物的染色体ＤＮＡ是否也能在此真核体系中进行核装配一直没有报道。我们以大肠杆菌染色体ＤＮＡ为材料，研究了它诱导的非细胞体系核装置。在光镜与电镜水平观察了核装配的过程。显微分光光度计扫描显示ＤＮＡ片段在核装配过程中经历了凝集－去凝集的变化。证明大肠杆菌染色体ＤＮＡ也

Nuclear reconstitution around purified E. coli DNA in cell-free extracts of Xenopus laevis eggs]

In recent years we have succeeded in inducing various exogenous DNA to assembly typical nuclei in cell-free Xenopus egg extracts. However, it is not clear if prokaryotic chromosomal DNA can also form the nucleus in this eukaryotic system. In our experiment, we used the purified E. coli. chromosomal DNA to reconstitute the nucleus in the nuclear reconstitution extracts and found that typical nuclei can also be assembled around added DNA. We observed the process of nuclear reconstitution by using light microscopy and electronic microscopy. The scanning results of chromatin-like materials and assembled nuclei by microspectrophotometer showed that E. coli, chromosal DNA underwent obvious change from condensation to decondensation during nuclear reconstitution. Incorporation of alpha-32P-dCTP into DNA suggested that the reconstituted nuclei were capable of DNA replication.