Efficient and sensitive assay for T-DNA-dependent transient gene expression |
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Authors: | L Rossi J Escudero B Hohn B Tinland |
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Institution: | (1) Friedrich Miescher-Institut, P.O. Box 2543, CH4002 Basel, Switzerland |
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Abstract: | We describe here a very sensitive and reproducible method to detect the efficiency ofAgrobacterium-mediated T-DNA transfer. This method is based on a quantitative assay of β-glucuronidase activity produced in the plant cell
upon transfer of T-DNA carrying a specialuidA gene construct. Analysis of the transfer efficiency of a transfer-proficient bacterium compared with that of the same bacterium
diluted at different ratios with a transfer-defective bacterium shows a high sensitivity of the β-glucuronidase activity in
the plant. Five orders of magnitude in T-DNA transfer efficiency can be covered when the activity is measured combining the
fluorimetric MUG assay (for high activity) and the histochemical X-Gluc assay (very sensitive for low activity). |
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Keywords: | β -glucuronidase transient expression T-DNA tobacco seedlings Nicotiana tabacum |
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