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