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Site-Directed φC31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria
JoVE Journal
Genética
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JoVE Journal Genética
Site-Directed φC31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria

Site-Directed φC31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria

DOI:
10.3791/62146-v

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09:38 min

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February 02, 2021

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, , ,
1Vector Biology Department,Liverpool School of Tropical Medicine, 2Department of Microbiology & Molecular Genetics,University of California, 3Department of Molecular Biology & Biochemistry,University of California

Capítulos

  • 00:04Introduction
  • 00:46Preparation of Plasmids for the Microinjection Mix
  • 02:33Microinjection of Embryos from an Anopheles Docking Line
  • 03:38Crossing and Screening of Transformed Individuals
  • 06:45Results: Validation of φC31 Single Integration and RMCE in An. stephensi Larvae
  • 08:56Conclusion

Summary

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The protocol describes how to achieve site-directed modifications in the genome of Anopheles malaria mosquitoes using the φC31 system. Modifications described include both the integration and the exchange of transgenic cassettes in the genome of attP-bearing docking lines.

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Keywords: Site-directed IntegrationAttB-mediated IntegrationCassette ExchangeAnophelesMalariaTransgenesisTransgenic MosquitoesEmbryo MicroinjectionMarker GeneHelper PlasmidIntegrase

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