Protocol for Production of a Genetic Cross of the Rodent Malaria Parasites

Sittiporn Pattaradilokrat; Jian Li; Xin-zhuan Su

doi:10.3791/2365

Protocol for Production of a Genetic Cross of the Rodent Malaria Parasites

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Protocol for Production of a Genetic Cross of the Rodent Malaria Parasites

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13:39 min

January 3, 2011

DOI: 10.3791/2365-v

Sittiporn Pattaradilokrat¹, Jian Li^1,2, Xin-zhuan Su¹

¹National Institute of Allergy and Infectious Diseases,National Institutes of Health, ²School of Life Science,Xiamen University

Overview

This video demonstrates the procedure for producing genetic crosses of the rodent malaria parasite Plasmodium yoelii to generate recombinant progeny. The method involves feeding mosquitoes with infected mice to facilitate genetic recombination.

Key Study Components

Area of Science

Malaria research
Genetic recombination
Parasitology

Background

Rodent malaria parasites can be genetically crossed to study their genetics.
Understanding genetic determinants is crucial for malaria research.
Recombinant progeny can provide insights into drug resistance and transmissibility.
This method can be applied to various rodent malaria species.

Purpose of Study

To produce genetic crosses of Plasmodium yoelii.
To generate recombinant progeny for further analysis.
To investigate genetic factors influencing malaria characteristics.

Methods Used

Injection of mice with a mixed inoculum of two parental clones.
Feeding infected mice to mosquitoes to facilitate zygote formation.
Harvesting sporozoites from mosquito salivary glands.
Re-injecting sporozoites into naive mice to obtain blood stage parasites.

Main Results

Successful generation of recombinant progeny from genetic crosses.
Characterization of recombinant clones through genotyping.
Identification of 5-10% of cloned lines as recombinants.
Insights into genetic determinants of drug resistance and transmissibility.

Conclusions

The method is effective for producing genetic crosses in malaria research.
Recombinant progeny can help answer critical questions in the field.
This approach can be adapted for other rodent malaria parasites.

Frequently Asked Questions

What is the significance of genetic crosses in malaria research?

Genetic crosses help identify genetic determinants of traits such as drug resistance and transmissibility.

How are recombinant progeny generated?

Recombinant progeny are generated by feeding mosquitoes with infected mice and harvesting sporozoites.

What percentage of cloned lines are typically recombinants?

Typically, 5-10% of cloned lines will be identified as recombinants.

Can this method be applied to other malaria species?

Yes, the method is applicable to various rodent malaria parasites.

What is the role of genotyping in this study?

Genotyping is used to characterize recombinant clones and identify genetic markers.

What are the main goals of this experimental procedure?

The main goals are to produce genetic crosses and generate recombinant progeny for research.

Genetic crosses of rodent malaria parasites are performed by feeding two genetically distinct parasites to mosquitoes. Recombinant progeny are cloned from mouse blood after allowing mosquitoes to bite infected mice. This video shows how to produce genetic crosses of Plasmodium yoelii and is applicable to other rodent malaria parasites.

The overall goal of this procedure is to produce a genetic cross of the rodent malaria parasite plasmodium yoi in order to generate recombinant progeny. This is achieved by first injecting a group of mice with a mixed inoculum containing the two parental clones of the parasites to be crossed. Infected mice are then used to feed mosquitoes inside, which the gat of each parental strains will fuse into a diploid zygote myotic division at this stage can lead to genetic recombination, thereby producing recombinant progeny.

The resulting sporozoites are then harvested from the mosquitoes sali glands and re-injected into naive recipient mice in order to obtain blood stage parasites among which are individual recombinant clones that can be isolated by limited dilution. In a successful experiment, five to 10%of the cloned lines will be recombinants, which can be characterized by genotyping for appropriate genetic markers. This method can help answer key question in the malaria research field, such as what are the genetic determinants of drug resistant ants and transmissibility in malaria?

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