Expression of plastocyanin and cytochrome f of the cyanobacterium Phormidium laminosum in Escherichia coli and Paracoccus denitrificans and the role of leader peptides

Citation
Bg. Schlarb et al., Expression of plastocyanin and cytochrome f of the cyanobacterium Phormidium laminosum in Escherichia coli and Paracoccus denitrificans and the role of leader peptides, GENE, 234(2), 1999, pp. 275-283
Citations number
33
Categorie Soggetti
Molecular Biology & Genetics
Journal title
GENE
ISSN journal
03781119 → ACNP
Volume
234
Issue
2
Year of publication
1999
Pages
275 - 283
Database
ISI
SICI code
0378-1119(19990708)234:2<275:EOPACF>2.0.ZU;2-U
Abstract
The gene for plastocyanin from the cyanobacterium Phormidium laminosum was successfully expressed in Escherichia coli. Expression of the gene for cyto chrome f resulted in the production of holocytochrome f in the periplasmic space of E. coli, but the yield was low. Expression in Paracoccus dentrific ans yielded no holoprotein. When the region encoding the cytochrome f leade r sequence was replaced with more typical bacterial leader sequences (those from the P. laminosum plastocyanin gene and the Paracoccus I versutus cyto chrome c-550 gene), much higher yields were consistently obtained in both s pecies. Overexpressed proteins were compared to those isolated from P. lami nosum and found to be identical in mass, isoelectric point, redox midpoint potential and (for plastocyanin) H-1-NMR spectrum. (C) 1999 Published by El sevier Science B.V. All rights reserved.