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Site-specific deletions in the tomato genome by the CinH-RS2 and ParA-MRS recombination systems

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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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Acknowledgments

We thank Professor Zhibiao Ye of Huazhong Agriculture University for technical support and Zhiguo Han of South China Botanical Garden for technical advice and discussion. This work was partially supported by Knowledge Innovation Project of The Chinese Academy of Sciences (KSCX2-EW-J-20), CAS/SAFEA International Partnership Program for Creative Research Teams Project, and the Chinese Ministry of Agriculture (2010ZX08010-001).

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  1. Yuan-Yeu Yau

    Present address: Department of Natural Sciences, Northeastern State University, Broken Arrow, OK, 74014, USA

Authors and Affiliations

  1. Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, 430074, China

    Yin Zhou & Ying Wang

  2. Plant Gene Expression Center, USDA/UC Berkeley, 800 Buchanan St., Albany, CA, 94710, USA

    Yuan-Yeu Yau & David W. Ow

  3. South China Botanical Garden, Xingke Road 723, Tianhe, Guangzhou, 510650, China

    David W. Ow

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  1. Yin Zhou
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  2. Yuan-Yeu Yau
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  4. Ying Wang
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Correspondence to David W. Ow or Ying Wang.

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Zhou, Y., Yau, YY., Ow, D.W. et al. Site-specific deletions in the tomato genome by the CinH-RS2 and ParA-MRS recombination systems. Plant Biotechnol Rep 6, 225–232 (2012). https://doi.org/10.1007/s11816-012-0217-4

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  • DOI: https://doi.org/10.1007/s11816-012-0217-4

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