Expression of a transgene exchanged by the recombinase-mediated cassette exchange (RMCE) method in plants |
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Authors: | Kazuya Nanto Kanna Sato Yoshihiro Katayama Hiroyasu Ebinuma |
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Institution: | (1) Forestly Science Laboratory, Nippon Paper Industries Co., Ltd, 5-21-1 Oji, Kita-ku, Tokyo 114-0002, Japan;(2) Institute of Symbiotic Science and Technology, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan;(3) R&D Division, Nippon Paper Industries Co., Ltd, 5-21-1 Oji, Kita-ku, Tokyo 114-0002, Japan |
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Abstract: | We developed a site-directed integration (SDI) system for Agrobacterium-mediated transformation to precisely integrate a single copy of a desired gene into a predefined target locus by recombinase-mediated
cassette exchange (RMCE). We produced site-specific transgenic tobacco plants from four target lines and examined expression
of the transgene in T1 site-specific transgenic tobacco plants, which were obtained by backcrossing. We found that site-specific
transgenic plants from the same target lines showed approximately the same level of expression of the transgene. Moreover,
we demonstrated that site-specific transgenic plants showed much less variability of transgene expression than random-integration
transgenic plants. Interestingly, transgenes in the same direction at the same target locus showed the same level of activity,
but transgenes in different directions showed different levels of activity. The expression levels of transgene did not correlate
with those of the target gene. Our results showed that the SDI system could benefit the precise comparisons between different
gene constructs, the characterization of different chromosomal regions and the cost-effective screening of reliable transgenic
plants. |
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Keywords: | Site-specific recombination Transgenic plant RMCE Gene expression |
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