Language:
English
In:
Molecular and biochemical parasitology, 2011, Vol.175(2), pp.205-208
Description:
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. ; Includes references ; p. 205-208.
Keywords:
Cell Cycle ; Trypanosoma Brucei ; Bsf ; Mrna Expression ; Pcf ; Quantitative Real Time Pcr ; Fluorescence-Activated Cell Sorting ; Qpcr ; Bloodstream Form ; Procyclic Form ; Facs
ISSN:
0166-6851
URL:
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