Alternatively spliced transcripts and novel pseudogenes of the Plasmodium falciparum resistance-associated locus pfcrt detected in East African malaria patients.
Objectives: Polymorphisms in the lysosomal transporter encoded by the pfcrt gene directly impact on Plasmodium falciparum susceptibility to aminoquinolines. The Lys76Thr mutation is the critical change conferring chloroquine resistance in vitro and in vivo, but always occurs with additional non-syno...
| Publicado en: | Journal of Antimicrobial Chemotherapy (JAC) Vol. 70; no. 1; pp. 116 - 124 |
|---|---|
| Autores principales: | , , , , , , , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
Oxford University Press / USA
Jan2015
|
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=109770255&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 109770255 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 03057453 N5O jtl: Journal of Antimicrobial Chemotherapy (JAC) issn: 03057453 maglogo: N pubinfo: dt: Jan2015 vid: 70 iid: 1 pid: 622 pub: Oxford University Press / USA artinfo: ui: 109770255 109770255 NLM25253286 2012838736 10.1093/jac/dku358 NLM25253286 PMC4267505 109770255 ppf: 116 ppct: 8 formats: tig: atl: Alternatively spliced transcripts and novel pseudogenes of the Plasmodium falciparum resistance-associated locus pfcrt detected in East African malaria patients. aug: au: Gadalla, Nahla B Malmberg, Maja Adam, Ishag Oguike, Mary C Beshir, Khalid Elzaki, Salah-Eldin Mukhtar, Izdihar Gadalla, Amal A Warhurst, David C Ngasala, Billy Mårtensson, Andreas El-Sayed, Badria B Gil, J Pedro Sutherland, Colin J sug: subj: RNA Malaria Membrane Transport Proteins Protozoa Proteins Genes RNA Metabolism Adult Amino Acids Child Child, Preschool DNA Female Infant Male Models, Biological Models, Theoretical Documentation Polymerase Chain Reaction Polymorphism, Genetic Molecular Structure Sequence Analysis Sudan Tanzania Funding Source Human Adult: 19-44 years Child: 6-12 years Child, Preschool: 2-5 years Infant: 1-23 months Female Male ab: Objectives: Polymorphisms in the lysosomal transporter encoded by the pfcrt gene directly impact on Plasmodium falciparum susceptibility to aminoquinolines. The Lys76Thr mutation is the critical change conferring chloroquine resistance in vitro and in vivo, but always occurs with additional non-synonymous changes in the pfcrt coding sequence. We sought to better describe pfcrt polymorphisms distal to codon 76.Methods: Small-volume samples (≤ 500 μL) of parasite-infected blood collected directly from malaria patients presenting for treatment in Sudan and Tanzania were immediately preserved for RNA extraction. The pfcrt locus was amplified from cDNA preparations by nested PCR and sequenced directly to derive full-length mRNA sequences.Results: In one of two sites in Sudan, two patients were found with an unorthodox spliced form of pfcrt mRNA in which two exons were skipped, but it was not possible to test for the presence of the putative protein products of these aberrant transcripts. Genomic DNA sequencing from dried blood spots collected in parallel confirmed the presence of spliced pfcrt pseudogenes in a minority of parasite isolates. Full-length cDNA from conventionally spliced mRNA molecules in all study sites demonstrated the existence of a variety of pfcrt haplotypes in East Africa, and thus provides evidence of intragenic recombination.Conclusions: The presence of pseudogenes, although unlikely to have any direct public health impact, may confound results obtained from simple genotyping methods that consider only codon 76 and the adjacent residues of pfcrt. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
|---|