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Laser capture microdissection of Plasmodium falciparum liver stages for mRNA analysis
- Source :
- Molecular and Biochemical Parasitology. 121:179-183
- Publication Year :
- 2002
- Publisher :
- Elsevier BV, 2002.
-
Abstract
- Plasmodium falciparum liver-stages are important targets for vaccine-induced protective immune responses and prophylactic treatment against malaria. Little is known of the gene expression profile of malaria parasites during their development inside hepatocytes. The sequencing of the P. falciparum genome and the development of DNA microarray technology give new opportunities to identify genes expressed during the development of Plasmodium. However, transcriptome analysis cannot currently be applied to the hepatic stages, due to difficulties in obtaining sufficient amounts of parasite material that lie among the large excess of host cell RNA. Here, we describe the isolation of liver-stages by a modified laser capture microdissection approach applied to human hepatocyte cultures infected with P. falciparum. RT-PCR amplification of several P. falciparum transcripts demonstrated the high quality of the RNA recovered after microdissection. This approach should enable analyses of P. falciparum transcriptome during its hepatic development and substantially assist the identification of new therapeutic and vaccine targets.
- Subjects :
- biology
Reverse Transcriptase Polymerase Chain Reaction
Lasers
Plasmodium falciparum
Protozoan Proteins
RNA
biology.organism_classification
Virology
Transcriptome
parasitic diseases
Gene expression
Hepatocytes
Animals
Humans
Parasitology
RNA, Messenger
DNA microarray
Molecular Biology
Gene
Cells, Cultured
RNA, Protozoan
Microdissection
Laser capture microdissection
Subjects
Details
- ISSN :
- 01666851
- Volume :
- 121
- Database :
- OpenAIRE
- Journal :
- Molecular and Biochemical Parasitology
- Accession number :
- edsair.doi.dedup.....e724746182e0f97a8da7a4085247d173