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Yeast As a Chassis for Developing Functional Assays to Study Human P53
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Yeast As a Chassis for Developing Functional Assays to Study Human P53

Yeast As a Chassis for Developing Functional Assays to Study Human P53

DOI:
10.3791/59071-v

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14:57 min

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August 04, 2019

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, , , , ,
1Mutagenesis and Cancer Prevention Unit,Ospedale Policlinico San Martino, 2Department CIBIO,University of Trento, 3LAQV/REQUIMTE, Laboratory of Microbiology, Faculty of Pharmacy,University of Porto

Chapters

  • 00:00Title
  • 00:54Construction of ADE2 or LUC1 Reporter Yeast Strain that Contains a Specific RE (yAFM-RE or yLFM-RE)
  • 03:06Evaluation of P53 Protein Transactivation Ability using the Qualitative Color-based ADE2 Yeast Assay
  • 04:27Evaluation of P53 Protein Transactivation Ability using the Quantitative Luminescence-based LUC1 Yeast Assay
  • 07:06Evaluation of P53 Protein Growth Inhibition using the Yeast Phenotypic Assay
  • 08:08Results
  • 13:51Conclusion

Summary

자동 번역

자동 번역

Presented here are four protocols to construct and exploit yeast Saccharomyces cerevisiae reporter strains to study human P53 transactivation potential, impacts of its various cancer-associated mutations, co-expressed interacting proteins, and the effects of specific small molecules.

Tags

YeastP53TranscriptionReporter GeneAdenine-2Firefly LuciferaseLithium AcetateTransformationPCRSanger SequencingTransactivationColor-based Assay

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