Site-specific deletions in the tomato genome by the CinH-<Emphasis Type="Italic">RS2</Emphasis> and ParA-<Emphasis Type="Italic">MRS</Emphasis> recombination systems |
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Authors: | Yin Zhou Yuan-Yeu Yau David W Ow Ying Wang |
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Institution: | 1.Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden,Chinese Academy of Sciences,Wuhan,China;2.Plant Gene Expression Center,USDA/UC Berkeley,Albany,USA;3.South China Botanical Garden,Guangzhou,China;4.Department of Natural Sciences,Northeastern State University,Broken Arrow,USA |
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Abstract: | We have tested the CinH-RS2 and ParA-MRS site-specific deletion systems in tomato (Solanum lycopersicum L.). The ParA-MRS system is derived from the broad-host-range plasmid RK2, where the 222 aa ParA recombinase recognizes a 133 bp multimer resolution
site (MRS). The CinH-RS2 system is derived from Acinetobacter plasmids pKLH2 and pKLH204, where the 188 amino acid CinH recombinase recognizes a 113-bp recombination site known as RS2. In this study, target lines containing a DNA segment flanked by recombination sites were crossed to recombinase-expressing
lines producing CinH or ParA recombinase. CinH-mediated recombination of RS2 substrates was detected in 2 of 3 F1 plants that harbor both the target and recombinase loci. On the other hand, recombination mediated by ParA was not detected
among F1 plants, but was found among 13 of 47 F2 plants. These data show that both systems can mediate site-specific DNA deletion in the tomato genome, and, upon further
refinement, can provide additional molecular tools for tomato improvement through precise genome manipulation. As the target
construct also contains additional recombination sites for site-specific integration by other recombination systems, these
tomato lines could be used for future testing of gene stacking through site-specific integration. |
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