Next generation sequencing analysis of transcriptomes of Plasmodium falciparum male and female gametocytes. Plasmodium falciparum

Investigation of overall expression level in Plasmodium falciparum male and female mature gametocytes, and detection of any transcriptional differences between male and female gametocytes. The Plasmodium falciparum parasite with green fluorescent protein (GFP) expression under the control of alpha tubulin II promoter facilitated the separation of male and female gametocyte. This engineered parasite strain in this study are further described in Miao J, Fan Q, Parker D, Li X, Li J, et al. (2013) Puf Mediates Translation Repression of Transmission-Blocking Vaccine Candidates in Malaria Parasites. PLoS Pathog 9(4): e1003268. doi: 10.1371/journal.ppat.1003268 Overall design: A 12 RNA-seq study was performed. Based on GFP intensity, equal number (4-5.5 X 10 6 ) of mature male and female gametocytes were separated by cell-sorter. Total RNA with and without rRNA depletion were processed for generation libraries using the Illumina TruSeq Stranded mRNA Library Preparation kit (Illumina Inc., San Diego, CA). Libraries were sequenced on an Illumina HiSeq 2500 in Rapid Run mode using 100 nucleotide single read sequencing. Altogether, this study investigated three experimental replicates of transcriptomes for mRNA and rRNA from male and female gametocytes (12 samples)