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In vitro reassembly of nuclear envelopes and organelles in xenopus egg extracts

Cell Research 2006年第7期16卷 632-640页

作者： Ping Lu~1 Hui Zheng~2 Zhonghe Zhai ~1College of Life Sciences ~2Institute of Molecular Medicine,Peking University Beijing 100871,China College of Life Sciences Peking University Beijing China Institute of Molecular Medicine Peking University Beijing China

We reconstituted bilayer nuclear membranes,multilayer membranes,and organelles from mixtures of xenopus laevis eggextracts and demembranated xenopus sperm *** proportions of the cytosolic and vesicular fractions from theeggs were used in the reconstitution mixtures.A cytosol:vesicle ratio of 10:1 promoted reassembly of the normal bilayernuclear membrane with inserted nuclear pore complexes around the decondensed xenopus sperm chromatin.A cytosol:vesicle ratio of 5:1 caused decondensed and dispersed sperm chromatin to be either surrounded by or divided by unusualmultilayer membrane structures with inlaid pore complexes.A cytosol:vesicle ratio of 2.5:1 promoted reconstitution ofmitochondria,endoplasmic reticulum networks,and Golgi *** reassembly of the endoplasmic reticulumand Golgi apparatus,vesicular fragments of the corresponding organelles fused together and changed their shape to formflattened cisternae,which were then stacked one on top of another.

关键词： cell-free system endoplasmic reticulum Golgi apparatus mitochondria nuclear reconstitution xenopus egg extracts

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Association of DNA with nuclear matrix in in vitro assembled nuclei induced by rDNA from Tetrahymena shanghaiensis in xenopus egg extracts

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Cell Research 1997年第1期7卷 107-117页

作者： CHEN YING BO ZHANG XIU FEN LI ZHONG HE ZHAI(Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871) Department of Cell Biology and Genetics College of Life Sciences Peking University Beijing

The nuclei assembled from exogenous DNA or chromatin in egg extracts resemble their in vivo counterparts in many aspects. However, the distribution pattern of DNA in these nuclei remains unknown. We introduced rDNA from the macronuclei of Tetrabymena into xenopus cellfree extracts to examine the association of specific DNA sequences with nuclear matrix (NM) in the nuclei assembled in vitro. Our previous works showed the 5'NTS (nontranscription sequences) of the rDNA specifically bind to the NM system in the macronuclei. We show now the rDNA could induce chromatin assembly and nuclear formation in xenopus cell-free system. When we extracted the NM system and compared the binding affinity of different regions of rDNA with the NM system, we found that the 5'NTS still hold their binding affinity with insoluble structure of the assembled nuclei in the extracts of xenopus eggs.

关键词： Nuclear assembly nuclear matrix xenopus egg extracts Tetrahymena rDNA

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