Please use this identifier to cite or link to this item:
https://doi.org/10.1111/tpj.12397
DC Field | Value | |
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dc.title | Direct visualization of Agrobacterium-delivered VirE2 in recipient cells | |
dc.contributor.author | Li, X. | |
dc.contributor.author | Yang, Q. | |
dc.contributor.author | Tu, H. | |
dc.contributor.author | Lim, Z. | |
dc.contributor.author | Pan, S.Q. | |
dc.date.accessioned | 2014-10-27T08:26:16Z | |
dc.date.available | 2014-10-27T08:26:16Z | |
dc.date.issued | 2014-02 | |
dc.identifier.citation | Li, X., Yang, Q., Tu, H., Lim, Z., Pan, S.Q. (2014-02). Direct visualization of Agrobacterium-delivered VirE2 in recipient cells. Plant Journal 77 (3) : 487-495. ScholarBank@NUS Repository. https://doi.org/10.1111/tpj.12397 | |
dc.identifier.issn | 09607412 | |
dc.identifier.uri | http://scholarbank.nus.edu.sg/handle/10635/100476 | |
dc.description.abstract | Agrobacterium tumefaciens is a natural genetic engineer widely used to deliver DNA into various recipients, including plant, yeast and fungal cells. The bacterium can transfer single-stranded DNA molecules (T-DNAs) and bacterial virulence proteins, including VirE2. However, neither the DNA nor the protein molecules have ever been directly visualized after the delivery. In this report, we adopted a split-GFP approach: the small GFP fragment (GFP11) was inserted into VirE2 at a permissive site to create the VirE2-GFP11 fusion, which was expressed in A. tumefaciens; and the large fragment (GFP1-10) was expressed in recipient cells. Upon delivery of VirE2-GFP11 into the recipient cells, GFP fluorescence signals were visualized. VirE2-GFP11 was functional like VirE2; the GFP fusion movement could indicate the trafficking of Agrobacterium-delivered VirE2. As the natural host, all plant cells seen under a microscope received the VirE2 protein in a leaf-infiltration assay; most of VirE2 moved at a speed of 1.3-3.1 μm sec-1 in a nearly linear direction, suggesting an active trafficking process. Inside plant cells, VirE2-GFP formed filamentous structures of different lengths, even in the absence of T-DNA. As a non-natural host recipient, 51% of yeast cells received VirE2, which did not move inside yeast. All plant cells seen under a microscope transiently expressed the Agrobacterium-delivered transgene, but only 0.2% yeast cells expressed the transgene. This indicates that Agrobacterium is a more efficient vector for protein delivery than T-DNA transformation for a non-natural host recipient: VirE2 trafficking is a limiting factor for the genetic transformation of a non-natural host recipient. The split-GFP approach could enable the real-time visualization of VirE2 trafficking inside recipient cells. © 2013 The Authors The Plant Journal © 2013 John Wiley & Sons Ltd. | |
dc.description.uri | http://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1111/tpj.12397 | |
dc.source | Scopus | |
dc.subject | Agrobacterium | |
dc.subject | Nicotiana benthamiana | |
dc.subject | protein delivery | |
dc.subject | Saccharomyces cerevisiae | |
dc.subject | T-DNA | |
dc.subject | technical advance | |
dc.subject | VirE2 | |
dc.subject | visualization | |
dc.type | Article | |
dc.contributor.department | BIOLOGICAL SCIENCES | |
dc.description.doi | 10.1111/tpj.12397 | |
dc.description.sourcetitle | Plant Journal | |
dc.description.volume | 77 | |
dc.description.issue | 3 | |
dc.description.page | 487-495 | |
dc.description.coden | PLJUE | |
dc.identifier.isiut | 000330079000013 | |
Appears in Collections: | Staff Publications |
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