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Adaption of the Leishmania Cell-Free Expression System to High-Throughput Analysis of Protein Interactions

Part of the Methods in Molecular Biology book series (MIMB,volume 2025)

Abstract

In this chapter, we present methods for adapting the eukaryotic cell-free expression system based on Leishmania tarentolae to high-throughput analysis of protein interactions. Specifically, we present a lysate optimization technique that minimizes the amount of unwanted premature termination products while balancing protein expression level and protein aggregation. Finally, we present methods for adapting the Leishmania cell-free system to the AlphaLISA-based protein interaction assay.

Key words

  • Leishmania tarentolae
  • mCherry
  • eGFP
  • High-throughput
  • Protein interactions
  • AlphaLISA

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  • DOI: 10.1007/978-1-4939-9624-7_19
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Correspondence to Kirill Alexandrov .

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Johnston, W.A., Moradi, S.V., Alexandrov, K. (2019). Adaption of the Leishmania Cell-Free Expression System to High-Throughput Analysis of Protein Interactions. In: Vincentelli, R. (eds) High-Throughput Protein Production and Purification. Methods in Molecular Biology, vol 2025. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9624-7_19

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  • DOI: https://doi.org/10.1007/978-1-4939-9624-7_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9623-0

  • Online ISBN: 978-1-4939-9624-7

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