lamblia an interesting system for studying eukaryotic evolution and the evolution of parasitism. Research on G. lamblia is aided by the fact that the entire life cycle
can be studied outside the host, and that the differentiation from cyst to trophozoite and the reverse process of encystation can be reproduced in vitro. Recently, the availability find more of the complete genome sequence [2–5] have facilitated genome-wide analyses. Although many Smoothened inhibitor Giardia proteins and organelles have been studied in detail, genome-wide studies of the transcriptome and proteome have been few [6–11]. No microarray analyses of the transcriptome of cysts obtained from infected animals have to our knowledge been performed. Serial Analysis of Gene Expression (SAGE) was used to survey changes in the G. lamblia transcriptome during encystation and excystation [9]. This study grouped about 10% of predicted G. lamblia genes into six clusters with related transcriptional profile. SAGE was also used to analyze the relative abundance of transcripts encoding cytoskeleton proteins [8]. This analysis found that the level of mRNA transcripts encoding proteins localized in the adhesive decreases as the parasite PKA activator encysts, and also found a lack of association between mRNA and protein level. Morf and co-workers focused on transcriptional changes associated with encystation [12]. This study used micoarrays to identify
genes which are induced during encystation and found evidence of transcriptional co-regulation mediated by a shared transcription factor binding motif in the promoter region of such genes. The extensive morphological changes which take place during the parasite’s life cycle have for years motivated the study of transcriptional regulation of selected genes during differentiation. Reverse-transcription
PCR has been frequently used to monitor changes in the level of specific mRNA transcripts, such as those encoding enzymes involved in energy metabolism [13], recombination [14], structural functions [15] or regulatory functions [16]. We wished to compare on a global level the transcriptional landscape of trophozoites and cysts. We found that in cysts many genes are either not transcribed, or that the transcripts they encode are too rare to Casein kinase 1 be detected with microarrays. Results Analysis of the cyst transcriptome The cyst and trophozoite transcriptome were compared by plotting mean Cy3 fluorescence values from six replicate microarrays hybridized with cDNA from independent live cyst suspensions and two replicate microarrays hybridized with trophozoite cDNA. The two trophozoites samples originated from a culture of assemblage B GS isolate in exponential phase of growth harvested 24 h post-inoculation and from a stationary culture harvested at 72 h. Cysts of assemblage B isolate H3 were obtained from experimentally infected gerbils. Their viability estimated by propidium iodide exclusion [17] ranged from 90% to 93% in three randomly selected cyst samples.