恶性疟原虫血液期转录位点和基因组位点的原位荧光可视化
In situ fluorescence visualization of transcription sites and genomic Loci in blood stages of Plasmodium falciparum.
作者信息
Mancio-Silva Liliana, Scherf Artur
机构信息
Unité de Biologie des Interactions Hôte-Parasite, Institut Pasteur, Paris, France.
出版信息
Methods Mol Biol. 2013;923:335-51. doi: 10.1007/978-1-62703-026-7_24.
Abstract
Fluorescence-based techniques have been used extensively in the malaria field to study the functional role of nuclear organization and gene positioning in blood stages of the human malaria parasite, Plasmodium falciparum. In this chapter, we present optimized protocols for bromouridine (BrUTP) incorporation into nascent RNA in live parasites and fluorescent in situ hybridization (FISH) in fixed parasites. Methodology to perform various combinations of the FISH assay, as well as a basic approach for quantitative analysis of nuclear position, is also described.
摘要
基于荧光的技术已在疟疾研究领域广泛应用，用于研究人类疟原虫恶性疟原虫血液阶段中核组织和基因定位的功能作用。在本章中，我们介绍了将溴尿苷（BrUTP）掺入活寄生虫新生RNA以及对固定寄生虫进行荧光原位杂交（FISH）的优化方案。还描述了进行FISH分析各种组合的方法以及核位置定量分析的基本方法。
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Proc Natl Acad Sci U S A. 2010 Aug 24;107(34):15117-22. doi: 10.1073/pnas.1001045107. Epub 2010 Aug 9.
2
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Mol Biochem Parasitol. 2010 Sep;173(1):48-52. doi: 10.1016/j.molbiopara.2010.04.006. Epub 2010 Apr 28.
3
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Nat Rev Genet. 2009 Jul;10(7):457-66. doi: 10.1038/nrg2592.
4
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Cell Host Microbe. 2009 Feb 19;5(2):179-90. doi: 10.1016/j.chom.2008.12.012.
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