A Modular Vector for Agrobacterium Mediated Transformation of Wheat |
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Authors: | Peters Noël R Ackerman Steven Davis Elizabeth A |
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Institution: | (1) Institute for Chemistry and Cell Biology, Harvard Medical School, 240 Longwood Ave., Boston, MA 02115, USA;(2) Biology Department, University of Massachusetts, Boston |
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Abstract: | Wheat (cv Chinese Spring) tissues were transformed using Agrobacterium tumefasciens and a new plasmid modular vector, pMVTBP. We constructed pMVTBP with unique restriction sites connecting (1) the CaMV 35S
promoter, (2) a Kozak sequence, (3) the FLAG epitope, (4) the (His)6 epitope, (5) a coding region (for wheat TATA Binding Protein, wTBP) and (6) the CaMV 35S 3′UTR. This vector thus allows easy
exchange of different regulatory or coding sequences. Explants of either germinating mature seeds, or immature embryos, were
induced to callus for up to two weeks, treated with virulence-induced bacteria for one hour, then regenerated into plantlets.
Transient expression of a GUS reporter gene, assayed at about one week, occurred in 10–12% of calluses. Expression of the
FLAG-tagged wTBP was also detected, by immunostaining. Stable expression, by selective growth on geneticin, and by GUS expression
at about six weeks, occurred in 1–2% of calluses, quite comparable to that achieved by other methods.
An erratum to this article is available at . |
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Keywords: | Agrobacterium modular vector transformation wheat |
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