Hybrid genes in the analysis of transformation conditions |
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Authors: | I. Negrutiu R. Shillito I. Potrykus G. Biasini F. Sala |
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Affiliation: | (1) Friedrich Miescher-Institut, P.O. Box 2543, CH-4002 Basel, Switzerland;(2) Dipartimento di genetica e microbiologia, Università di Pavia, Via S. Epifanio, 14-27100 Pavia, Italy;(3) Present address: Instituut voor Moleculaire Biologie, 65 Paardenstraat, B-1640 Sint-Genesius-Rode, Belgium |
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Abstract: | ![]() Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×105 treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the competence for transformation has a species-specific component.Abbreviations PEG polyethyleneglycol - RTF relative transformation frequency - ATF absolute transformation frequency |
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Keywords: | transformation MgCl2-PEG-electroporation competence protoplasts |
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