Author | Bourgard, Catarina | |
Author | Lopes, Stefanie C. P. | |
Author | Lacerda, Marcus V. G. | |
Author | Albrecht, Letusa | |
Author | Costa, Fabio T. M. | |
Access date | 2023-02-07T14:18:41Z | |
Available date | 2023-02-07T14:18:41Z | |
Document date | 2021 | |
Citation | BOURGARD, Catarina et al. A suitable RNA preparation methodology for whole transcriptome shotgun sequencing harvested from Plasmodium vivax‑infected patients. Scientific Reports, [s. l.], v. 11, n. 5089, p. 1-12, 2021. | en_US |
ISSN | 2045-2322 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/56919 | |
Sponsorship | Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) e Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). | en_US |
Language | eng | en_US |
Rights | open access | en_US |
Title | A suitable RNA preparation methodology for whole transcriptome shotgun sequencing harvested from Plasmodium vivax‑infected patients | en_US |
Type | Article | en_US |
DOI | 10.1038/s41598-021-84607-w | |
Abstract | Plasmodium vivax is a world-threatening human malaria parasite, whose biology remains elusive. The unavailability of in vitro culture, and the difficulties in getting a high number of pure parasites makes RNA isolation in quantity and quality a challenge. Here, a methodological outline for RNA-seq from P. vivax isolates with low parasitemia is presented, combining parasite maturation and enrichment with efficient RNA extraction, yielding ~ 100 pg.µL−1 of RNA, suitable for SMART-Seq Ultra-Low Input RNA library and Illumina sequencing. Unbiased coding transcriptome of ~ 4 M reads was achieved for four patient isolates with ~ 51% of transcripts mapped to the P. vivax P01 reference genome, presenting heterogeneous profiles of expression among individual isolates. Amongst the most transcribed genes in all isolates, a parasite-staged mixed repertoire of conserved parasite metabolic, membrane and exported proteins was observed. Still, a quarter of transcribed genes remain functionally uncharacterized. In parallel, a P. falciparum Brazilian isolate was also analyzed and 57% of its transcripts mapped against IT genome. Comparison of transcriptomes of the two species revealed a common trophozoite-staged expression profile, with several homologous genes being expressed. Collectively, these results will positively impact vivax research improving knowledge of P. vivax biology. | en_US |
Affilliation | University of Campinas. Institute of Biology. Department of Genetics, Evolution, Microbiology and Immunology. Prof. Dr. Luiz Jacintho da Silva. Laboratory of Tropical Diseases. Campinas, SP, Brazil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto Leônidas & Maria Deane. Manaus, AM, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto Leônidas & Maria Deane. Manaus, AM, Brasil. | en_US |
Affilliation | University of Campinas. Institute of Biology. Department of Genetics, Evolution, Microbiology and Immunology. Prof. Dr. Luiz Jacintho da Silva. Laboratory of Tropical Diseases. Campinas, SP, Brazil. | en_US |
Affilliation | University of Campinas. Institute of Biology. Department of Genetics, Evolution, Microbiology and Immunology. Prof. Dr. Luiz Jacintho da Silva. Laboratory of Tropical Diseases. Campinas, SP, Brazil. | en_US |
Subject | Plasmodium vivax | en_US |
Subject | Malaria | en_US |