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无标记基因删除由絮凝血盒等位基因交换突变在衣原体沙眼
Journal JoVE
Immunologie et infection
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Journal JoVE Immunologie et infection
Markerless Gene Deletion by Floxed Cassette Allelic Exchange Mutagenesis in Chlamydia trachomatis
DOI:

10:35 min

January 30, 2020

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Chapitres

  • 00:04Introduction
  • 00:27Design and Assembly of pSUmC-4.0 with Homology Arms Specific to the Gene of Interest
  • 02:20Transformation of C. trachomatis with pSUmC 4.0 + Homology Arms for Gene Deletion by Allelic Exchange
  • 05:54Clonal Isolation of Green-only C. trachomatis Deletion Mutant Containing the loxP Flanked Selection Cassette by Limiting Dilution
  • 07:38Transformation of C. trachomatis FRAEM Mutant with pSU-Cre to Initiate Removal of loxP Flanked Selection Cassette
  • 08:42Results: FLAEM Enables Markerless Gene Deletion in C. trachomatis and alleviates Cassette-induced Polar Effects
  • 09:47Conclusion

Summary

Traduction automatique

本文描述的是使用絮状盒状体等位基因突变(FLAEM)在衣原体沙眼中进行靶向、无标记基因删除的方法。

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