title: Production of a recombinant antimicrobial peptide in transgenic plants using a modified VMA intein expression system.
abstract: Tobacco plants were engineered to express SMAP-29, a mammalian antimicrobial peptide of innate immunity, as fusion protein with modified vacuolar membrane ATPase intein. The peptide was purified taking advantage of the intein-mediated self-cleaving mechanism. SMAP-29 was immunologically detected in the chromatographic eluate and appeared tightly bound to copurified plant proteins. Electrophoretic separation under disaggregating conditions indicated that the recombinant peptide was cleaved off by intein at the expected site and an overlay gel assay demonstrated that the peptide retained antimicrobial activity. These results indicate that a modified intein expression system can be used to produce pharmaceutical peptides in transgenic plants.
authors.affiliation: DPVTA, Università di Udine, Via delle Scienze 208, Udine, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Morassutti
authors.forename: Francesca
authors.initials: F
authors.lastname: De Amicis
authors.forename: Barbara
authors.initials: B
authors.lastname: Skerlavaj
authors.forename: Margherita
authors.initials: M
authors.lastname: Zanetti
authors.forename: Stefano
authors.initials: S
authors.lastname: Marchetti
mesh_terms: D001798:Blood Proteins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D054804:Cathelicidins; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry / Q000378:metabolism*; D019076:Transgenes; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D054804:Cathelicidins; D011993:Recombinant Fusion Proteins; C403305:SMAP29 protein, Ovis aries; D025262:Vacuolar Proton-Translocating ATPases