[Nuclear reconstitution around purified HeLa chromosomes in a cell-free system from Xenopus eggs]

Shi Yan Sheng Wu Xue Bao. 1994 Mar;27(1):51-9.
[Article in Chinese]

Abstract

Activated Xenopus egg extracts are capable of reconstituting nucleus around the added DNA or chromatin. To understand better the nuclear reconstitution process and the structure of the assembled nucleus, we have developed a cell-free system using activated Xenopus egg extracts and metaphase chromosomes of HeLa cells. We demonstrate that the extracts are efficient promoting the decondensation of the metaphase chromosomes and the nuclear reconstitution around the chromosomes, and that the nuclear reconstitution is a precise process. The reassembled nucleus possesses a typical nuclear envelope, pore complexes, nuclear lamina, nuclear matrix, chromatin, etc., as shown by electron microscopy, embedment and embedment-free electron microscopy, and immunofluorescence method. We also found that the annulate lamellae directly take part in nuclear envelope and pore complex reconstitution by fusing with the double membrane, having reassembled on the surface of chromatin/chromosomes.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus* / ultrastructure
  • Cell-Free System
  • Chromosomes*
  • Female
  • HeLa Cells / cytology
  • Humans
  • Metaphase
  • Ovum*
  • Tissue Extracts
  • Xenopus laevis

Substances

  • Tissue Extracts