Linked Life Data

17697981

Source:http://linkedlifedata.com/resource/pubmed/id/17697981

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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2007-8-16
pubmed:abstractText
We developed a new series of binary vectors useful for Gateway cloning to facilitate transgenic experiments in plant biotechnology. The new system, Gateway Binary Vectors (pGWBs) realized efficient cloning, constitutive expression using the cauliflower mosaic virus (CaMV) 35S promoter and the construction of fusion genes by simple clonase reaction with an entry clone. The reporters employable in this system are beta-glucuronidase (GUS), synthetic green fluorescent protein with S65T mutation (sGFP), luciferase (LUC), enhanced yellow fluorescent protein (EYFP), and enhanced cyan fluorescent protein (ECFP). The tags available are 6xHis, FLAG, 3xHA, 4xMyc, 10xMyc, GST, T7-epitope, and tandem affinity purification (TAP). In total, 13 kinds of reporter or tag were arranged and were almost applicable to both N- and C-fusions. The pGWBs could be used for many purposes, such as promoter::reporter analysis, observation of subcellular localization by the expression of proteins fused to a reporter or tag, and analysis of protein-protein interaction by copurification and immunodetection experiments. The pGWBs were constructed with modified pBI101 containing a CaMV35S promoter-driven hygromycin phosphotransferase (HPT) gene as the second selection marker. We also constructed pGWBs with the marker HPT driven by the nopaline synthase promoter. By using the pGWB system, the expression of tagged proteins, and the localization of GFP-fused proteins were easily analyzed. Moreover, tissue-specific and inducible gene expression using a promoter was also monitored with pGWBs. It is expected that, the pGWB system will serve as a powerful tool for plasmid construction in plant research.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
1389-1723
pubmed:author
pubmed:issnType
Print
pubmed:volume
104
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
34-41
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:17697981-Amino Acid Sequence, pubmed-meshheading:17697981-Artificial Gene Fusion, pubmed-meshheading:17697981-Base Sequence, pubmed-meshheading:17697981-Caulimovirus, pubmed-meshheading:17697981-Cell Line, pubmed-meshheading:17697981-Cloning, Molecular, pubmed-meshheading:17697981-Gene Fusion, pubmed-meshheading:17697981-Genes, Reporter, pubmed-meshheading:17697981-Genetic Vectors, pubmed-meshheading:17697981-Glucuronidase, pubmed-meshheading:17697981-Green Fluorescent Proteins, pubmed-meshheading:17697981-Molecular Sequence Data, pubmed-meshheading:17697981-Phosphotransferases (Alcohol Group Acceptor), pubmed-meshheading:17697981-Plants, pubmed-meshheading:17697981-Promoter Regions, Genetic, pubmed-meshheading:17697981-Recombinant Fusion Proteins, pubmed-meshheading:17697981-Transformation, Genetic
pubmed:year
2007
pubmed:articleTitle
Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation.
pubmed:affiliation
Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University, 1060 Nishikawatsu, Matsue, Japan. tnakagaw@life.shimane-u.ac.jp
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't