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Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae
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JoVE Journal Genetica
Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae

Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae

DOI:
10.3791/62131-v

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06:29 min

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April 15, 2021

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, , , , , ,
1Department of Vector Biology,Liverpool School of Tropical Medicine, 2IMBB FORTH, 3Department of Biology,University of Crete

Capitoli

  • 00:05Introduction
  • 00:49Collection of Pupae for Microscopic Characterization and Identification of Fluorescent Markers
  • 01:45Sexing Pupae
  • 02:25Setting Up GAL4-UAS (Upstream Activation Sequences) System Crosses and Inducing Egg Laying
  • 03:20Embryo Fixation and Bleaching
  • 04:15Results: Analysis of Anopheles gambiae Pupae Expressing Fluorescent Markers Driven by 3xP3 Promoter, Tissue-specific Expression Visualization, and Embryo Clearing
  • 05:53Conclusion

Summary

Traduzione automatica

Traduzione automatica

The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation.

Tags

GAL4-UAS SystemAnopheles GambiaeFunctional GeneticsGenetic ModificationGene ExpressionPhenotypic CharacterizationInsecticide ResistancePathogen TransmissionGenetic ControlPupae CollectionFluorescent MarkerExternal GenitaliaSexingAdult TransferBlood FeedingOviposition

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