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具有eGFP组成型表达的 利什曼原虫 菌株的发展
JoVE Journal
Genetics
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JoVE Journal Genetics
Development of Leishmania Species Strains with Constitutive Expression of eGFP
DOI:

10:03 min

April 21, 2023

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Chapters

  • 00:04Introduction
  • 00:27Amplification and Insertion of the eGFP Gene into the pLEXSY‐2.1 Vector
  • 02:39Linearization of the pLEXSY Expression Plasmid
  • 03:26Transfection of L. panamensis and L. donovani by Electroporation
  • 05:00Polyclonal Selection in Culture
  • 06:41Confirmation of Genomic Integration
  • 07:09Cloning by Limiting Dilution
  • 08:12Results: Confirmation of eGFP Cassette Expression and Chromosomal Insertion
  • 09:40Conclusion

Summary

Automatic Translation

在这里,我们描述了用于生成使用pLEXSY系统将eGFP基因表达为稳定整合转基因的L. panamensisL. donovani 菌株的方法。通过限制稀释来克隆转染的寄生虫,并选择两种物种中荧光强度最高的克隆进一步用于药物筛选测定。

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