Periplasmic fractionation of escherichia coli yields recombinant plastocyanin despite the absence of a signal sequence

Joel A. Ybe, Michael H. Hecht

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Poplar plastocyanin has been expressed in E. coli from a synthetic gene cloned into the T7 expression system. Despite the absence of a signal sequence, large quantities of the recombinant protein were readily obtained by procedures typically used to isolate proteins from the bacterial periplasm. Several different fractionation methods were equally successful. The presence of plastocyanin in these fractions does not reflect wholesale leakage of intracellular proteins, since neither β-galactosidase activity nor the bulk of Escherichia coli proteins were released by the fractionation. The identity of the overexpressed protein was unequivocally proven to be poplar plastocyanin by N-terminal amino acid sequence analysis and by spectroscopic characterization of the purified blue copper protein.

Original languageEnglish (US)
Pages (from-to)317-323
Number of pages7
JournalProtein Expression and Purification
Volume5
Issue number4
DOIs
StatePublished - Jan 1 1994

All Science Journal Classification (ASJC) codes

  • Biotechnology

Fingerprint Dive into the research topics of 'Periplasmic fractionation of escherichia coli yields recombinant plastocyanin despite the absence of a signal sequence'. Together they form a unique fingerprint.

  • Cite this