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Review
. 2007 Dec;145(4):1118-28.
doi: 10.1104/pp.107.106104.

Delivery of multiple transgenes to plant cells

Mery Dafny-Yelin  1 , Tzvi Tzfira
Affiliations
Review

Delivery of multiple transgenes to plant cells

Mery Dafny-Yelin et al. Plant Physiol. 2007 Dec.
No abstract available

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Figures

Figure 1.
Figure 1.
Methods for the assembly of multiple-gene binary plasmids. A, The Cre/loxP-mediated multigene assembly process. i, Cre/loxP recombination of the pYLVS-A plasmid into the pYLTAC747 acceptor binary plasmid. ii, Release of the pYLVS backbone by I-SceI digestion and ligation with a LS linker. This ligation abolishes the I-SceI site from pYLTAC747-A. iii, Cre/loxP recombination of the pYLSV-B plasmid into the pYLTAC747-A and release of the pYLSV backbone by PI-SceI digestion and ligation with a LV linker (iv). B, The MultiRound Gateway assembly process. i, Gateway recombination between attL1 and attR1 and between attL1 and attR1 sites by LR clonase and conversion of the ccdB-based binary pDEST vector into a sacB-based Destination vector. ii, Gateway recombination between attL3 and attR3 and between attL4 and attR4 sites by LR clonase and reconversion of the destination binary vector into a ccdB-based plasmid. C, The homing endonucleases pRCS/pAUX vector system. Assembly of a multigene binary plasmid is achieved by successive cloning of various gene expression cassettes using rare-cutting homing endonucleases.
Figure 2.
Figure 2.
The general structure of a pSAT-based plant expression vector. A typical vector is composed of promoter and terminator sequences, flanked by AgeI and NcoI and XbaI and NotI sites, respectively. This arrangement allows for easy replacement of these regulatory elements in nearly all of the pSAT-based vectors as well as the construction of various specialized tags and functional constructs between the NcoI and XbaI sites. Fully assembled expression cassettes can be mobilized between different pSAT backbones by AgeI-NotI and be cloned into pRCS-based vectors using various rare cutters. [See online article for color version of this figure.]
See this image and copyright information in PMC

References

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