实验动物总则

Atomic Force Microscopy (AFM) is a powerful tool for exploring the interaction between ligands and receptors, as well as their exact locations on the red cell surface. Here we discuss current and future applications for AFM based single-molecule force spectroscopy to study adhesion of Plas ...

The altered deformability of erythrocytes infected with Plasmodium falciparum is central in malaria �pathogenesis, as it influences the hemodynamic properties of the infected cell and its retention in the spleen. Exported parasite proteins, as well as the shape and volume of the paras ...

An increased level of cytosolic free calcium (Ca2+) is an essential second messenger for apical organelle discharge in Plasmodium falciparum merozoites. Here, we describe a method for isolation of viable and invasive P. falciparum merozoites. We also describe methods to measure cytoso ...

Cellular imaging has reemerged in recent years as a powerful approach to provide researchers with a direct measure of essential molecular events in a cell’s life, ranging in scale from broad morphological observations of whole cells to intricate single molecule imaging. When combined wi ...

Metabolomics is an increasingly common analytical approach for investigating metabolic networks of pathogenic organisms. This may be of particular use in the study of parasitic infections due to the intrinsic metabolic connection between the parasite and its host. In vitro cultures ...

The central role played by protein phosphorylation in the regulation of eukaryotic cellular processes calls for detailed investigations of this phenomenon in malaria parasites. Here, we describe protocols to measure the activity of protein kinases (using either recombinant pro ...

Transcriptome analysis by next-generation sequencing (RNA-seq) allows investigation of a transcriptome at unsurpassed resolution. One major benefit is that RNA-seq is independent of a priori knowledge on the sequence under investigation, thereby also allowing analysis of poo ...

The application of DNA microarray technologies to malaria genomics has been widely used but has been limited by sample availability and technical variability. To address these issues, we present a microarray hybridization protocol that has been optimized for use with two new Agilent Tec ...

DNA microarray is presently one of the most powerful and fastest growing technologies for genomic research of infectious diseases. Accordingly, DNA microarray-based global analyses of Plasmodium parasites provided many insights into the general biology of malaria infection. F ...

Real-time polymerase chain reaction (PCR), or quantitative PCR (qPCR), is a rapid, sensitive, and specific method used for a broad variety of applications including quantitative gene expression analysis, DNA copy number measurement, characterization of gene and chromosomal dele ...

Genetically modified Plasmodium parasites are central gene function reagents in malaria research. The Rodent Malaria genetically modified DataBase (RMgmDB) (www.pberghei.eu) is a manually curated Web - based repository that contains information on genetically modified r ...

DNA of Plasmodium berghei is difficult to manipulate in Escherichia coli by conventional restriction and ligation methods due to its high content of adenine and thymine (AT) nucleotides. This limits our ability to clone large genes and to generate complex vectors for modifying the parasite ...

Gene manipulation is an invaluable tool to investigate and understand the biology of an organism. Although this technology has been applied to both the human and rodent malarial parasites (RMP), Plasmodium berghei in particular offers a more robust system due to a higher and more efficient tr ...

Genetic manipulation of Plasmodium falciparum remains very challenging, mainly due to the parasite genome’s high A/T-richness and low transfection efficiency. This chapter includes methods for generating transient and stable transfections by electroporation, allelic re ...

Anopheles gambiae mosquitoes are the major vectors of human malaria parasites. However, mosquitoes are not passive hosts for parasites, actively limiting their development in vivo. Our current understanding of the mosquito antiparasitic response is mostly based on the phenotypic ...

Here, we describe the methodology for transient transfection of Plasmodium vivax. The ability to genetically manipulate P. vivax has rendered this important human malaria parasite more amenable to molecular investigation. However, a systematic analysis of this parasite and its di ...

We provide a series of protocols that have been used for the cyclic transmission of rodent malaria parasites in the laboratory. This is now possible both in vivo and in vitro. We focus on the least “resource intensive” and generic methods that we find applicable to any parasite–host combination. Non ...

Long-term in vitro cultures of blood-stage parasites are so far feasible only for Plasmodium falciparum and P. knowlesi. In this chapter, we describe short-term ex vivo culturing of P. cynomolgi and P. vivax. We also describe long-term in vitro culturing of P. knowlesi as well as some techniques for sy ...

The ookinete is the motile form of the malaria parasite that invades the mosquito midgut epithelium to initiate sporogony. Differentiation of ingested gametocytes into ookinetes in the mosquito midgut lumen and the subsequent interaction with the luminal surface of the midgut epith ...

Production of gametocytes in vitro is essential for studies of Plasmodium falciparum sexual stages. Here, we describe procedures for the high-yield production and fractionation of P. falciparum gametocytes stages I to V.