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It was show11 that nuclear reassembly was induced by small pieces of DNA fragments in cell-free extracts ofXenopus. In an attempt to learn the relationship between the nuclear reassembly and nucleosome/chromatin assembly, limited amounts
of CM-Cellulose are used to eliminate the capacity of the egg extract S-150 to assemble chromatin. while the forming of nucleosomes
is checked with DNA supercoiling by plasmid DNA pBR322 incubated in the extract, and further analysed by micrococcal nuclease
digestion. This depleted extract is then used to induce nuclear reassembly around demembraned sperms with membrane vesicles.
It is found that CM-Cellulose depletes histones H2A and H2B efficiently and blocks the assembly of nucleosomes, the demembraned
sperms are yet reconstituted into nuclei in the treated S-150, although the chromatin in reassembled nuclei does not produce
protected DNA fragments when digested with micrococcal nuclease. It suggests that in the cell-free system ofXenopus, DNA can be formed into nuclei without assembly of nucleosomes or chromatin.
Projrrt supported by the National Natural Science Foundation of China (Grant No. 39730240) 相似文献
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Gurdon JB 《Journal of biosciences》2005,30(1):11-14
When nuclei of somatic cells are transplanted to enucleated eggs ofXenopus, a complete reprogramming of nuclear function can take place. To identify mechanisms of nuclear reprogramming, somatic nuclei
can be transplanted to growing meiotic oocytes ofXenopus, and stem cell genes activated without DNA replication. The combination of somatic cell nuclear transfer with morphogen signalling
and the community effect may lead towards the possibility of cell replacement therapy. When mechanisms of nuclear reprogramming
are understood, it may eventually be possible to directly reprogramme human somatic cell nuclei without the use of eggs. 相似文献
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Transcriptional activation of Xenopus class III genes in chromatin isolated from sperm and somatic nuclei 总被引:8,自引:0,他引:8
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A P Wolffe 《Nucleic acids research》1989,17(2):767-780