High Throughput Protein Expression and Purification pp 105-115

Part of the Methods in Molecular Biology book series (MIMB, volume 498)

A Family of LIC Vectors for High-Throughput Cloning and Purification of Proteins

  • William H. Eschenfeldt
  • Stols Lucy
  • Cynthia Sanville Millard
  • Andrzej Joachimiak
  • I. Donnelly Mark

Summary

Fifteen related ligation-independent cloning vectors were constructed for high-throughput cloning and purification of proteins. The vectors encode a TEV protease site for removal of tags that facilitate pro tein purification (his-tag) or improve solubility (MBP, GST). Specialized vectors allow coexpression and copurification of interacting proteins, or in vivo removal of MBP by TVMV protease to improve screening and purification. All target genes and vectors are processed by the same protocols, which we describe here.

Key words:

Structural genomics High throughput Protein purification Ligation-independ ent cloning Coexpression In vivo proteolysis Maltose-binding protein TEV protease TVMV protease 

Copyright information

© Humana Press, a part of Springer Science + Business Media 2009

Authors and Affiliations

  • William H. Eschenfeldt
    • 1
  • Stols Lucy
    • 1
  • Cynthia Sanville Millard
    • 1
  • Andrzej Joachimiak
    • 1
  • I. Donnelly Mark
    • 1
  1. 1.Biosciences Division, Argonne National LaboratoryLemontUSA

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