Expression of functional Raphanus sativus antifungal protein in yeast.
Rs-AFP2 is a 51 amino acid cysteine-rich peptide isolated from radish (Raphanus sativus) seeds that exhibits potent inhibitory activity against filamentous fungi. A cDNA clone encoding the Rs-AFP2 preprotein was modified by recombinant DNA methods to allow expression in the yeast Saccharomyces cerevisiae. This peptide was expressed in yeast as a fusion protein carrying at its N-terminus the prepro-sequences derived from the precursor of the yeast pheromone mating factor alpha 1. These sequences allow secretion of the biologically active peptide in a correctly processed form. Deletion of the mating factor alpha 1 pro-peptide drastically reduced the expression level of the peptide.
Alves
A L
AL
F.A. Janssens Laboratory of Genetics, Catholic University of Leuven, Heverlee, Belgium.
De Samblanx
G W
GW
Terras
F R
FR
Cammue
B P
BP
Broekaert
W F
WF
D000595:Amino Acid Sequence; D000935:Antifungal Agents; D001483:Base Sequence; D005670:Fusarium / Q000187:drug effects; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012639:Seeds / Q000737:chemistry*; D014675:Vegetables
D000935:Antifungal Agents; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins