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Expression and Purification of Soluble Adenylyl Cyclase from Escherichia coli

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G Protein Signaling

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 237))

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Summary

This chapter outlines methods to purify soluble adenylyl cyclase (AC7) expressed in an Escherichia coli (E. coli) heterologous expression system. Guidelines are provided for constructing the expression plasmids, optimizing expression, culturing, and purifying the proteins. Purification requires two chromatographic steps. A histidine tag (H6) is incorporated into the expression vector and utilized for affinity purification on a Ni-NTA column. Subsequently, an anion exchange column is employed to further purify the protein.

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Author information

Authors and Affiliations

  1. Ben May Institute for Cancer Research and Committee on Neurobiology, University of Chicago, Chicago, IL

    Jeff A. Beeler & Wei-Jen Tang

Authors
  1. Jeff A. Beeler
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  2. Wei-Jen Tang
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Editor information

Editors and Affiliations

  1. Department of Pharmacology and Physiology, University of Rochester School of Medicine, Rochester, NY

    Alan V. Smrcka

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© 2004 Humana Press Inc.

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Beeler, J.A., Tang, WJ. (2004). Expression and Purification of Soluble Adenylyl Cyclase from Escherichia coli . In: Smrcka, A.V. (eds) G Protein Signaling. Methods in Molecular Biology™, vol 237. Humana Press. https://doi.org/10.1385/1-59259-430-1:39

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  • DOI: https://doi.org/10.1385/1-59259-430-1:39

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-137-0

  • Online ISBN: 978-1-59259-430-6

  • eBook Packages: Springer Protocols

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