| Reliable quantification of cell cycle-dependent mRNA abundance using fluorescence-activated cell sorting in Trypanosoma brucei. | |
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MedLine Citation:
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PMID: 20933544 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
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Very little is known about cell cycle-dependent regulation of mRNA in Trypanosoma brucei, the causative agent of African sleeping sickness. Methods to synchronize cell cycle progression are inefficient or subject the parasites to non-physiological conditions and stress. We developed a fluorescence-activated cell sorting-based method to analyze steady-state mRNA levels in individual cell cycle phases. Normalization of the data was the most challenging problem because internal standards for cell cycle-regulated genes are not available for trypanosomes. Hence, we introduced an external standard (so-called "spike") to compensate for technically derived variations in processing cells and RNA samples. Validation of this method with a limited number of genes unraveled a transient up-regulation during S and G2/M phases for all mRNAs analyzed. |
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Authors:
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Ferdinand Bucerius; Markus Kador; Michael Boshart; Christian J Janzen |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-10-08 |
Journal Detail:
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Title: Molecular and biochemical parasitology Volume: 175 ISSN: 1872-9428 ISO Abbreviation: Mol. Biochem. Parasitol. Publication Date: 2011 Feb |
Date Detail:
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Created Date: 2010-12-20 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8006324 Medline TA: Mol Biochem Parasitol Country: Netherlands |
Other Details:
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Languages: eng Pagination: 205-8 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 Elsevier B.V. All rights reserved. |
Affiliation:
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University of Munich (LMU), Department Biology I, Genetics, Martinsried, Germany. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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