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位点导向 的φC31介导的疟疾按蚊载体中的整合和盒式交换

JoVE Journal
Genetics

This content is Free Access.

JoVE Journal Genetics

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

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位点导向的φC31介导的疟疾按蚊载体中的整合和盒式交换

DOI:

10.3791/62146-v

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09:38 min

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February 02, 2021

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Adriana Adolfi², Amy Lynd, Gareth J. Lycett, Anthony A. James³

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

Chapters

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

Automatic Translation

该协议描述了如何使用φC31系统在按蚊的基因组中实现位点导向的修饰。所描述的修饰包括在携带attP的对接线的基因组中转基因盒的整合和交换。

Tags

Keywords: Site-directed Integration AttB-mediated Integration Cassette Exchange Anopheles Malaria Transgenesis Transgenic Mosquitoes Embryo Microinjection Marker Gene Helper Plasmid Integrase

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