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Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin
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Biologia
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JoVE Journal Biologia
Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin

Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin

DOI:
10.3791/3562-v

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07:56 min

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January 17, 2012

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, ,
1Department of Orthopaedic Surgery,NYU Hospital for Joint Diseases, 2Department of Cell Biology,New York University School of Medicine

Capitoli

  • 00:05Titolo
  • 01:07Small Scale Transformation of Bait Plasmid
  • 02:12Bait Validation
  • 03:09cDNA Library Screening
  • 05:06X-gal Assay
  • 05:59Retransformation in the MY2H Assay
  • 06:47Isolation of Progranulin-Interacting Proteins
  • 07:32Conclusion

Summary

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We have modified the conventional yeast two-hybrid screening, an effective genetic tool in identifying protein interaction. This modification markedly shortens the process, reduces the workload, and most importantly, reduces the number of false positives. In addition, this approach is reproducible and reliable.

Tags

Modified Yeast-two-hybrid SystemProteinsInteractingGrowth FactorProgranulinPGRNGranulin Epithelin PrecursorAutocrine Growth FactorPhysiologic ProcessesDisease ProcessesInflammationHost DefenseNeurotrophic FactorPGRN GeneFrontotemporal DementiaCartilage Development And DegradationBinding Receptor(s)Modified Yeast Two-hybrid ApproachGAL4 Based 2-hybrid SystemFalse Positive ClonesReproducible And Reliable System

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