title: Visualisation of plastids in endosperm, pollen and roots of transgenic wheat expressing modified GFP fused to transit peptides from wheat SSU RubisCO, rice FtsZ and maize ferredoxin III proteins.
abstract: The ability to target marker proteins to specific subcellular compartments is a powerful research tool to study the structure and development of organelles. Here transit sequences from nuclear-encoded, plastid proteins, namely rice FtsZ, maize non-photosynthetic ferredoxin III (FdIII) and the small subunit of RubisCO were used to target a modified synthetic GFP (S65G, S72A) to plastids. The localisations of the fusion proteins expressed in transgenic wheat plants and under the control of the rice actin promoter were compared to an untargeted GFP control. GFP fluorescence was localised to non-green plastids in pollen, roots and seed endosperm and detected in isolated leaf chloroplasts using a GFP-specific antibody. Transit peptides appeared to influence the relative fluorescence intensities of plastids in different tissues. This is consistent with differential targeting and/or turnover of GFP fusion proteins in different plastid types. Replacement of GFP sequences with alternative coding regions enables immediate applications of our vectors for academic research and commercial applications.
authors: Rothamsted Research, Harpenden, Hertfordshire, UK.
authors: Lucia F
authors: LF
authors: Primavesi
authors: Huixia
authors: H
authors: Wu
authors: Elisabeth A
authors: EA
authors: Mudd
authors: Anil
authors: A
authors: Day
authors: Huw D
authors: HD
authors: Jones
mesh_terms: D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins; D001483:Base Sequence; D015153:Blotting, Western; D005288:Ferredoxins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D015388:Organelles / Q000235:genetics / Q000378:metabolism; D012275:Oryza / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D018087:Plastids / Q000235:genetics / Q000378:metabolism*; D011058:Pollen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics / Q000378:metabolism*; D003313:Zea mays / Q000235:genetics / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; C095113:FTSZ protein, Arabidopsis; D005288:Ferredoxins; D010446:Peptide Fragments; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012273:Ribulose-Bisphosphate Carboxylase
references: Annu Rev Cell Dev Biol. 1996;12:1-26
references: 8970720
references: Plant Physiol. 1989 Apr;89(4):1193-7
references: 16666683
references: Bioinformatics. 2002 Feb;18(2):298-305
references: 11847077
references: Protein Sci. 1999 May;8(5):978-84
references: 10338008
references: Curr Opin Plant Biol. 2003 Jun;6(3):215-22
references: 12753970
references: Physiol Plant. 2002 May;115(1):144-154
references: 12010478
references: Plant Cell Rep. 2003 Mar;21(7):659-68
references: 12789416
references: J Biol Chem. 1996 Dec 6;271(49):31227-33
references: 8940125
references: J Exp Bot. 2001 Apr;52(356):529-39
references: 11373302
references: Transgenic Res. 1996 May;5(3):213-8
references: 8673150
references: J Exp Bot. 2001 Apr;52(357):857-63
references: 11413223
references: J Exp Bot. 2005 Mar;56(413):787-97
references: 15699062
references: Plant Physiol. 1991 May;96(1):77-83
references: 16668188
references: Plant J. 2001 Aug;27(3):257-65
references: 11532171
references: Plant Cell Rep. 2003 Jan;21(5):467-74
references: 12789450
references: Plant Sci. 2000 May 15;154(1):89-98
references: 10725562
references: J Exp Bot. 2000 Aug;51(349):1357-61
references: 10944148
references: Nature. 2002 Nov 21;420(6913):316-20
references: 12447439
references: Plant Physiol. 2001 Apr;125(4):2154-63
references: 11299394
references: Curr Biol. 1996 Mar 1;6(3):325-30
references: 8805250
references: Transgenic Res. 2004 Dec;13(6):523-30
references: 15672833
references: Plant Physiol. 2004 Nov;136(3):3457-66
references: 15489278
references: Annu Rev Plant Physiol Plant Mol Biol. 2000 Jun;51:111-140
references: 15012188
references: J Exp Bot. 2003 May;54(386):1321-33
references: 12709478
references: FEBS Lett. 2006 Nov 27;580(27):6509-12
references: 17101134
references: Nature. 1995 Feb 23;373(6516):663-4
references: 7854443
references: Gene. 1996;173(1 Spec No):33-8
references: 8707053
references: J Cell Sci. 1989 Jul;93 ( Pt 3):491-500
references: 2606940
references: Plant Physiol. 1991 Sep;97(1):375-80
references: 16668395
references: Plant Mol Biol. 1998 Sep;38(1-2):191-207
references: 9738967
references: J Cell Biol. 2000 Nov 13;151(4):945-50
references: 11076976
references: Biochem Biophys Res Commun. 2001 Sep 21;287(2):462-7
references: 11554751
references: J Exp Bot. 2001 Apr;52(357):865-74
references: 11413224
references: Gene. 2003 Nov 27;320:97-108
references: 14597393
references: Transgenic Res. 2008 Jun;17(3):425-36
references: 17638109
references: Plant J. 2004 Aug;39(4):655-67
references: 15272881
references: Plant Cell. 1993 Oct;5(10):1383-99
references: 8281040
references: Plant Cell. 1998 Dec;10(12):1991-2004
references: 9836740
references: Mol Genet Genomics. 2001 Mar;265(1):161-71
references: 11370863
references: Trends Plant Sci. 1999 Jul;4(7):253-257
references: 10407440
references: Mol Gen Genet. 1991 Dec;231(1):150-60
references: 1753941
references: J Cell Biol. 2001 Apr 2;153(1):111-20
references: 11285278
references: Science. 1997 Jun 27;276(5321):2039-42
references: 9197266
references: Methods Enzymol. 1986;118:253-70
references: 2419734