Generation of Arabidopsis Mutants by Heterologous Expression of a Full-Length cDNA Library from Tomato Fruits |
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Authors: | Yi-Hong Wang Chris M Mosebach Abraham S Kibbey Marcie K Ryhal Angelica D Jones Julie A Palmer Leon V Kochian |
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Institution: | (1) School of Science, Penn State University, Behrend College, 4205 College Dr., Erie, PA 16563, USA;(2) US Plant Soil and Nutrition Lab, Cornell University, Ithaca, NY 14853, USA;(3) Present address: Hamilton Hall, University of Louisiana at Lafayette, P.O. Box 44650, Lafayette, LA 70504-4650, USA |
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Abstract: | Heterologous expression of cDNA library in Arabidopsis and other plants has been used for gene identifications. To identify
functions of tomato genes, we expressed a tomato full-length cDNA library in Arabidopsis thaliana and generated over 7,000 mutants. We constructed a tomato cDNA library with a plant transformation-ready binary vector that
contained a higher percentage of full-length cDNAs since synthesized double-stranded cDNA was size-selected using gel electrophoresis,
with cDNA sizes of 2–5 kb being gel-purified for ligation onto the binary vector. Sequencing of 81 cDNA clones indicates that
75% (61) are full-length genes, which is similar to sequencing of inserted cDNA in Arabidopsis. The library was used to transform
Arabidopsis plants. Among the 7,000 mutants, one was found to be a dwarf due to the expression of an ATP synthase, and another
vegetative mutant did not produce flowers even after 7 months. The technique was validated by reintroducing the tomato ribosomal
protein L9 gene and can be used in any other plant species as a gene discovery tool. |
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