实验动物总则

Infections with human malaria parasites in New World monkeys offer opportunities to determine host-parasite interactions and relationships, to produce malarial antigens for diagnostic and molecular characterization, to conduct drug and vaccine efficacy trials, and to prod ...

The liver stages of the malaria parasite have long been a difficult part of the life cycle to study. Very little is known about the localization of the parasite within the liver and no method for the estimation of liver parasite burden is in common use. Differences in liver parasite burden would provide a go ...

Microsatellites (MS) are simple sequence repeats (SSRs) such as (CA)n, (TAA)n, or (TA)n that have been found in all eukaryotes studied (1-3). The number of repeated nucleotides per unit is generally limited to between 1 and 5, in contrast to the longer repeats characteristic of minisatellites. Whe ...

A standard method for evaluating inhibitory effects on preerythrocytic parasites in murine malaria models is to measure the time to patency of a blood-stage infection following a sporozoite challenge. The prevention of a blood-stage infection or a delay in patency suggests that preery ...

This chapter describes methods relating to the handling and processing of Anopheles mosquitoes captured during malaria vector field studies. Areas covered include the following methods

It is now established that Plasmodium falciparum parasites found in any one area are highly diverse and that individual hosts, vertebrate and insect, are often concurrently infected by multiple parasite lines (for example, see refs. 1-4). Different parasite lines of the same Plasmodium sp ...

The isolation of high-quality genomic DNA is often the initial step for molecular genetic analyses and crucial to many applications within the molecular research of malaria. Criteria for quality of DNA are purity, stability, and integrity and size of the molecules. Quantity also often is an im ...

The isolation of RNA and its subsequent characterization by various applications, such as the generation of cDNA libraries, Northern blot analysis, and reverse transcriptase polymerase chain reaction studies (RT-PCR), are fundamental for understanding the mechanisms underl ...

Southern blotting, first described by E. M. Southern in 1975 (1), is one of the cornerstones in molecular biology. The idea to immobilize DNA on a solid support was first proposed by Denhardt (2). Based on this, methods were developed for the identification of specific sequences in dot blots and recombi ...

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting are complementary methods for separating and detecting the presence of a specific protein from a complex mixture. Proteins, from a cell extract, for example, are separated electrophor ...

In the natural and untreated intermediate hosts, Plasmodium infections are maintained for extended and often lifelong periods. Parasitemia generally reaches its highest level in the first peak after inoculation, and only rises thereafter during increasingly brief and intersp ...

To achieve a more complete understanding of malaria parasite biology and its interaction with its host, it is essential to be able to study the transcription and expression of parasite genes. Northern blot analysis and RNase protection assays are commonly used to study gene expression (1). How ...

The genetic identification of malaria parasites has become an important task for epidemiological studies and for laboratory-based studies (1–7). A large number of adapted field isolates have been cultured and widely circulated during the past years. However, only few of these have been cl ...

The technique of in situ RNA hybridization provides a means of examining RNA expression within individual parasites at many stages of development in both the vertebrate and mosquito hosts. The protocols described in this chapter have been developed with the aim of preserving the morpholo ...

Sequencing of the entire genome from the human malaria parasite, Plasmodium falciparum, began in earnest in 1996 with the formation of an international consortium of scientists and funding agencies (1). Due to the instability of the highly adenine (A) and thymine (T)-rich P. falciparum DNA in la ...

Genomic DNA libraries represent the total complement of the genetic information of an organism’s DNA, as opposed to cDNA libraries, which contain only the protein encoding sequences expressed at a particular stage of the life cycle. Ideally, a genomic library contains the coding and control ...

Mung bean nuclease (MBN) has been used typically for its single-stranded nuclease activity (1). Under defined reaction conditions, however, which include altered solvation and elevated temperature, hypersensitive sites surrounding coding regions become sensitive to nucle ...

cDNA libraries represent the genetic information encoded in the messenger RNA (mRNA) of a particular tissue or organism. Construction of a cDNA library begins with the isolation of purified and full-length RNA. This isolation is made difficult by the fact that RNA molecules are exceptional ...

Subtractive hybridization techniques are designed to deplete one cDNA population (the “target”) of sequences common to a second group of cDNAs (the “driver”), thereby effectively enriching the target population for unique sequences. The applications of this powerful methodology ...

The discovery by Burke et al. in 1987 (1) that yeast can accept large pieces of heterologous DNA as yeast artificial chromosomes (YAC) marked a milestone in the analysis of complex genomes. While standard prokaryotic cloning systems, such as plasmids and cosmids, are limited by the size of the DNA frag ...