Expression of the Arabidopsis thaliana AtJ2 cochaperone protein in Pichia pastoris

Rengang Zhou, Barbara Kroczyńska, Jan A. Miernyk*

*Corresponding author for this work

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

A vector was constructed for intracellular expression of the Arabidopsis thaliana DnaJ homologue AtJ2 in the methylotrophic yeast Pichia pastoris. The vector includes DNA encoding an amino-terminal histidine-tag, to simplify protein purification. Shake-flask cultures could be induced to produce approximately 250 mg/L of AtJ2. Purified recombinant AtJ2 was able to stimulate the ATPase activities of both the Escherichia coli and Zea mays cytoplasmic Stress70 chaperone proteins five- to ninefold. The carboxy terminus of AtJ2 is -CAQQ, a protein farnesylation motif. When transformed P. pastoris was induced to synthesize AtJ2 in the presence of [3H]mevalonolactone, radioactivity was incorporated into the protein, suggesting farnesylation. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)253-258
Number of pages6
JournalProtein Expression and Purification
Volume19
Issue number2
DOIs
StatePublished - Jan 1 2000

ASJC Scopus subject areas

  • Biotechnology

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