Expression and affinity purification of recombinant proteins from plants

Urvee A. Desai, Gargi Sur, Sylvia Daunert, Ruth Babbitt, Qingshun Li

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

With recent advances in plant biotechnology, transgenic plants have been targeted as an inexpensive means for the mass production of proteins for biopharmaceutical and industrial uses. However, the current plant purification techniques lack a generally applicable, economic, large-scale strategy. In this study, we demonstrate the purification of a model protein, β-glucuronidase (GUS), by employing the protein calmodulin (CaM) as an affinity tag. In the proposed system, CaM is fused to GUS. In the presence of calcium, the calmodulin fusion protein binds specifically to a phenothiazine-modified surface of an affinity column. When calcium is removed with a complexing agent, e.g., EDTA, calmodulin undergoes a conformational change allowing the dissociation of the calmodulin-phenothiazine complex and, therefore, permitting the elution of the GUS-CaM fusion protein. The advantages of this approach are the fast, efficient, and economical isolation of the target protein under mild elution conditions, thus preserving the activity of the target protein. Two types of transformation methods were used in this study, namely, the Agrobacterium-mediated system and the viral-vector-mediated transformation system.

Original languageEnglish (US)
Pages (from-to)195-202
Number of pages8
JournalProtein Expression and Purification
Volume25
Issue number1
DOIs
StatePublished - Jan 1 2002
Externally publishedYes

Keywords

  • Affinity tag
  • Calmodulin
  • Expression
  • Isolation
  • Transgenic plants

ASJC Scopus subject areas

  • Biochemistry

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