Oleosin di-or tri-meric fusions with GFP undergo correct targeting and provide advantages for recombinant protein production

dc.contributor.author	Banilas, G	en
dc.contributor.author	Daras, G	en
dc.contributor.author	Rigas, S	en
dc.contributor.author	Moloney, MM	en
dc.contributor.author	Hatzopoulos, P	en
dc.date.accessioned	2014-06-06T06:51:24Z
dc.date.available	2014-06-06T06:51:24Z
dc.date.issued	2011	en
dc.identifier.issn	09819428	en
dc.identifier.uri	http://dx.doi.org/10.1016/j.plaphy.2010.12.002	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/5491
dc.subject	Fusion expression	en
dc.subject	Homo-oligomers	en
dc.subject	Oilbody stability	en
dc.subject	Oilbody targeting	en
dc.subject	Oleosins	en
dc.subject	Recombinant proteins	en
dc.subject.other	Arabidopsis protein	en
dc.subject.other	green fluorescent protein	en
dc.subject.other	oleosin protein, Arabidopsis	en
dc.subject.other	recombinant protein	en
dc.subject.other	Arabidopsis	en
dc.subject.other	article	en
dc.subject.other	fluorescence microscopy	en
dc.subject.other	genetics	en
dc.subject.other	metabolism	en
dc.subject.other	polymerase chain reaction	en
dc.subject.other	transgenic plant	en
dc.subject.other	Arabidopsis	en
dc.subject.other	Arabidopsis Proteins	en
dc.subject.other	Green Fluorescent Proteins	en
dc.subject.other	Microscopy, Fluorescence	en
dc.subject.other	Plants, Genetically Modified	en
dc.subject.other	Polymerase Chain Reaction	en
dc.subject.other	Recombinant Proteins	en
dc.subject.other	Arabidopsis	en
dc.title	Oleosin di-or tri-meric fusions with GFP undergo correct targeting and provide advantages for recombinant protein production	en
heal.type	journalArticle	en
heal.identifier.primary	10.1016/j.plaphy.2010.12.002	en
heal.publicationDate	2011	en
heal.abstract	Plant oleosins are small proteins embedded within the phospholipid monolayer separating the triacylglycerol storage site of embryo-located oilbodies from the cytoplasm of oilseed cells. The potential of oleosins to act as carriers for recombinant proteins foreign to plant cells has been well established. Using this approach, the recombinant polypeptide is accumulated in oilbodies as a fusion with oleosin. DNA constructs having tandemly arranged oleosins followed by GFP or flanked by oleosins were used to transform Arabidopsis plants. In all cases the green fluorescence revealed that the fusion polypeptide had a native conformation and the recombinant proteins were correctly targeted to seed oilbodies. Mobilization of lipids was not retarded when using homo-dimer or -trimer oleosin fusions, since seed production, germination rates and seedling establishment were similar among all constructs, and comparable to wild-type Arabidopsis plants. Plant physiology and growth of recombinant lines were similar to wild-type plants. The construct specifying two oleosins flanking the GFP polypeptide revealed interesting properties regarding both the accumulation and the relative stability of the oilbody protein assembly. Although expression levels varied among transgenic lines, those transgenes accumulated significantly higher levels of fusion proteins as compared to previously reported values obtained by a single-oleosin configuration, reaching up to 2.3% of the total embryo proteins. These results shows that the expression cassettes comprising three oleosin molecules in frame to the GFP molecule or two oleosins flanking the GFP could be advantageous over the single-oleosin configuration for higher production and better commercialization of this plant biotechnological platform without jeopardizing plant vigour and physiology or oilbody stability. © 2010 Elsevier Masson SAS.	en
heal.journalName	Plant Physiology and Biochemistry	en
dc.identifier.issue	2	en
dc.identifier.volume	49	en
dc.identifier.doi	10.1016/j.plaphy.2010.12.002	en
dc.identifier.spage	216	en
dc.identifier.epage	222	en