RRC ID 39581
Author Masuta C, Yamana T, Tacahashi Y, Uyeda I, Sato M, Ueda S, Matsumura T.
Title Development of clover yellow vein virus as an efficient, stable gene-expression system for legume species.
Journal Plant J
Abstract A highly infectious cDNA clone of clover yellow vein virus (pClYVV) was tested as a viral vector, especially for legume species. The genes for green fluorescent protein (GFP) and soybean glutamine synthetase (GS) were inserted between the genes for P1 and HC-Pro on pClYVV to create three recombinant plasmids: pClYVV-GFP, pClYVV-GFP-GS, and pClYVV-GFP:GS. In the former two constructs all the junctions between the inserted proteins contained the sequences of protease cleavage recognition sites, whereas the third construct expressed a fusion of GFP and GS. Western blot analyses showed that GFP and GS appeared to have been precisely excised from the viral polyprotein with the viral proteases (P1 and NIa). Under UV irradiation, green fluorescence was detected in infected broad bean, kidney bean, and soybean plants. The stability of the constructs in the symptomatic tissues was confirmed by RT-PCR and Western blot analyses. The plants expressing GS together with GFP became tolerant to the herbicide glufosinate, and flowered early. As the GS gene, one of the nodulin genes for nitrogen fixation, is expressed in legume species, this system will be useful for examining the function of genes important to legume plants.
Volume 23(4)
Pages 539-46
Published 2000-8-1
DOI 10.1046/j.1365-313x.2000.00795.x
PII tpj795
PMID 10972880
MeSH Blotting, Western Fabaceae / genetics* Fabaceae / metabolism Fabaceae / virology Gene Expression* Gene Transfer Techniques* Genes, Plant* Genetic Vectors Glutamate-Ammonia Ligase / genetics Green Fluorescent Proteins Luminescent Proteins / genetics Plants, Medicinal* Potyvirus / genetics* Potyvirus / metabolism Reverse Transcriptase Polymerase Chain Reaction
IF 6.141
Times Cited 56
DNA material pCIYVV-Pst/CP (RDB08246)