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从爪蟾卵母细胞制备灯刷染色体铺片的技巧和窍门。

Tips and tricks for preparing lampbrush chromosome spreads from Xenopus tropicalis oocytes.

机构信息

Institut Jacques Monod, UMR7592, CNRS and University Paris Diderot-Paris 7, Department of Molecular and Cellular Pathology, Paris, France.

出版信息

Methods. 2010 May;51(1):37-44. doi: 10.1016/j.ymeth.2010.01.014. Epub 2010 Jan 18.

Abstract

Due to their large size and fine organization, lampbrush chromosomes (LBCs) of amphibian oocytes have been for decades one of the favorite tools of biologists for the analysis of transcriptional and post-transcriptional processes at the cytological level. The emergence of the diploid Xenopus tropicalis amphibian as a model organism for vertebrate developmental genetics and the accumulation of sequence data made available by its recent genomic sequencing, strongly revive the interest of LBCs as a powerful tool to study genes expressed during oogenesis. We describe here a detailed protocol for preparing LBCs from X. tropicalis oocyte and give practical advice to encourage a large number of researchers to become familiar with these chromosomes.

摘要

由于两栖类卵母细胞的灯刷染色体(LBCs)体积较大、结构精细,几十年来一直是生物学家分析转录和转录后过程的首选工具之一。二倍体非洲爪蟾(Xenopus tropicalis)作为脊椎动物发育遗传学的模式生物的出现,以及其最近基因组测序所提供的序列数据的积累,强烈地重新激发了 LBCs 作为研究卵母细胞发生过程中表达基因的有力工具的兴趣。我们在这里描述了一种从 X. tropicalis 卵母细胞中制备 LBCs 的详细方案,并提供了实用建议,以鼓励更多的研究人员熟悉这些染色体。

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