Cre recombinase-mediated site-specific recombination
between plant chromosomes. |
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Authors: | M Qin C Bayley T Stockton D W Ow |
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Affiliation: | Plant Gene Expression Center, U.S. Department of Agriculture, Albany, CA 94710. |
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Abstract: | We report the use of the bacteriophage P1 Cre-loxsystem for generating conservative site-specific recombination between tobaccochromosomes. Two constructs, one containing a promoterless hygromycin-resistancegene preceded by a lox site (lox-hpt) and the other containing a cauliflowermosaic virus 35S promoter linked to a lox sequence and the cre coding region(35S-lox-cre), were introduced separately into tobacco plants. Crosses betweenplants harboring either construct produced plants with the two constructssituated on different chromosomes. Plants with recombination events wereidentified by selecting for hygromycin resistance, a phenotype expressed uponrecombination. Molecular analysis showed that these recombination eventsoccurred specifically at the lox sites and resulted in the reciprocal exchangeof flanking host DNA. Progenies of these plants showed 67-100% cotransmission ofthe new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferentialcosegregation of translocated chromosomes. These results illustrate thatsite-specific recombination systems can be useful tools for the large-scalemanipulation of eukaryotic chromosomes in vivo. |
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