Production of Xenopus tropicalis egg extracts to identify microtubule-associated RNAs.
Production of Xenopus tropicalis egg extracts to identify microtubule-associated RNAs.
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DOI：
10.3791/50434
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发表时间：
2013-06-27
期刊：
影响因子：
--
通讯作者：
Blower MD
中科院分区：
文献类型：
--
作者：
Sharp JA;Blower MD
Many organisms localize mRNAs to specific subcellular destinations to spatially and temporally control gene expression. Recent studies have demonstrated that the majority of the transcriptome is localized to a nonrandom position in cells and embryos. One approach to identify localized mRNAs is to biochemically purify a cellular structure of interest and to identify all associated transcripts. Using recently developed high-throughput sequencing technologies it is now straightforward to identify all RNAs associated with a subcellular structure. To facilitate transcript identification it is necessary to work with an organism with a fully sequenced genome. One attractive system for the biochemical purification of subcellular structures are egg extracts produced from the frog Xenopus laevis. However, X. laevis currently does not have a fully sequenced genome, which hampers transcript identification. In this article we describe a method to produce egg extracts from a related frog, X. tropicalis, that has a fully sequenced genome. We provide details for microtubule polymerization, purification and transcript isolation. While this article describes a specific method for identification of microtubule-associated transcripts, we believe that it will be easily applied to other subcellular structures and will provide a powerful method for identification of localized RNAs.
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影响因子：
11.4
作者：
Harland, Richard M.;Grainger, Robert M.
通讯作者：
Grainger, Robert M.
影响因子：
7.8
作者：
Brown, Katherine S.;Blower, Michael D.;Heald, Rebecca
通讯作者：
Heald, Rebecca
影响因子：
4
作者：
Mingle, LA;Okuhama, NN;Liu, G
通讯作者：
Liu, G
DOI：
10.1083/jcb.200705163
发表时间：
2007-12-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Blower MD;Feric E;Weis K;Heald R
通讯作者：
Heald R
影响因子：
64.5
作者：
Lecuyer, Eric;Yoshida, Hideki;Krause, Henry M.
通讯作者：
Krause, Henry M.