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使用基于路西法抹去的生长测定，确定化学抑制剂对细胞内弓形体贡迪生长的抑制效率
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Immunology and Infection
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JoVE Journal Immunology and Infection
Determination of Chemical Inhibitor Efficiency against Intracellular Toxoplasma Gondii Growth Using a Luciferase-Based Growth Assay
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使用基于路西法抹去的生长测定，确定化学抑制剂对细胞内弓形体贡迪生长的抑制效率

DOI:
10.3791/60985-v

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09:49 min

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April 29, 2020

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, , , , ,
1Department of Biological Sciences,Clemson University, 2Eukaryotic Pathogens Innovation Center,Clemson University

Chapters

  • 00:04Introduction
  • 00:49Luciferase-Based ToxoplasmaGrowth Assay
  • 02:25Chemical Compound Inhibition Efficacy Evaluation
  • 04:08CRISPR-Cas9-Based Gene Deletion
  • 05:59Toxoplasma Transfection
  • 06:51Knockout Parasite Cloning
  • 08:06Results: Representative LHVS Inhibition Efficacy Against Intracellular Toxoplasma Growth
  • 09:22Conclusion

Summary

Automatic Translation

Automatic Translation

本文介绍的是一种协议，用于使用基于荧光酶的生长测定来评估化合物对弓形体淋酸体内生长的抑制作用。该技术用于通过基因删除相应的靶基因来确认抑制特异性。以LHVS对TgCPL蛋白酶的抑制为例。

Tags

Toxoplasma GondiiLuciferase-based Growth AssayChemical InhibitorDrug TargetCRISPR-Cas9Intracellular PathogenHigh-throughput ScreeningParasite Doubling TimeHuman Foreskin FibroblastsLuciferase SubstrateMicroplate ReaderNormalized Fold-changeLinear Regression

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