pGD vectors: versatile tools for the expression of green and red fluorescent protein fusions in agroinfiltrated plant leaves

Goodin, MM, Dietzgen, RG, Schichnes, D, Ruzin, S and Jackson, AO (2002) pGD vectors: versatile tools for the expression of green and red fluorescent protein fusions in agroinfiltrated plant leaves. Plant Journal, 31 3: 375-383. doi:10.1046/j.1365-313X.2002.01360.x


Author Goodin, MM
Dietzgen, RG
Schichnes, D
Ruzin, S
Jackson, AO
Title pGD vectors: versatile tools for the expression of green and red fluorescent protein fusions in agroinfiltrated plant leaves
Journal name Plant Journal   Check publisher's open access policy
ISSN 0960-7412
Publication date 2002-08-01
Year available 2002
Sub-type Article (original research)
DOI 10.1046/j.1365-313X.2002.01360.x
Open Access Status DOI
Volume 31
Issue 3
Start page 375
End page 383
Total pages 9
Place of publication OXFORD
Publisher BLACKWELL PUBLISHING LTD
Language eng
Abstract We have constructed a matched set of binary vectors designated pGD, pGDG and pGDR for the expression and co-localization of native proteins and GFP or DsRed fusions in large numbers of plant cells. The utility of these vectors following agroinfiltration into leaves has been demonstrated with four genes from Sonchus yellow net virus , a plant nucleorhabdovirus, and with a nucleolar marker protein. Of the three SYNV proteins tested, sc4 gave identical localization patterns at the cell wall and nucleus when fused to GFP or DsRed. However, some differences in expression patterns were observed depending on whether DsRed or GFP was the fusion partner. In this regard, the DsRed:P fusion showed a similar pattern of localization to GFP:P, but localized foci appeared in the nucleus and near the periphery of the nucleus. Nevertheless, the viral nucleocapsid protein, expressed as a GFP:N fusion, co-localized with DsRed:P in a subnuclear locale in agreement with our previous observations (Goodin et al ., 2001). This locale appears to be distinct from the nucleolus as indicated by co-expression of the N protein, DsRed:P and a nucleolar marker AtFib1 fused to GFP. The SYNV M protein, which is believed to be particularly prone to oligomerization, was detectable only as a GFP fusion. Our results indicate that agroinfiltration with bacteria containing the pGD vectors is extremely useful for transient expression of several proteins in a high proportion of the cells of Nicotiana benthamiana leaves. The GFP and DsRed elements incorporated into the pGD system should greatly increase the ease of visualizing co-localization and interactions of proteins in a variety of experimental dicotyledonous hosts.
Keyword protein expression plasmids
DsRed
red fluorescent protein
Gfp
agroinfiltration
transient gene expression
Yellow Net Virus
Transgenic Plants
Gene
Rhabdovirus
Polymerase
Coral
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads - Archived
 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 200 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 210 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Mon, 10 Oct 2011, 18:56:28 EST by System User