Abstract
We report the use of the bacteriophage P1 Cre-lox system for generating conservative site-specific recombination between tobacco chromosomes. Two constructs, one containing a promoterless hygromycin-resistance gene preceded by a lox site (lox-hpt) and the other containing a cauliflower mosaic virus 35S promoter linked to a lox sequence and the cre coding region (35S-lox-cre), were introduced separately into tobacco plants. Crosses between plants harboring either construct produced plants with the two constructs situated on different chromosomes. Plants with recombination events were identified by selecting for hygromycin resistance, a phenotype expressed upon recombination. Molecular analysis showed that these recombination events occurred specifically at the lox sites and resulted in the reciprocal exchange of flanking host DNA. Progenies of these plants showed 67-100% cotransmission of the new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferential cosegregation of translocated chromosomes. These results illustrate that site-specific recombination systems can be useful tools for the large-scale manipulation of eukaryotic chromosomes in vivo.
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- A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
- Bayley C. C., Morgan M., Dale E. C., Ow D. W. Exchange of gene activity in transgenic plants catalyzed by the Cre-lox site-specific recombination system. Plant Mol Biol. 1992 Jan;18(2):353–361. doi: 10.1007/BF00034962. [DOI] [PubMed] [Google Scholar]
- Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chou T. B., Perrimon N. Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila. Genetics. 1992 Jul;131(3):643–653. doi: 10.1093/genetics/131.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clausen R E, Cameron D R. Inheritance in Nicotiana Tabacum. Xviii. Monosomic Analysis. Genetics. 1944 Sep;29(5):447–477. doi: 10.1093/genetics/29.5.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dale E. C., Ow D. W. Gene transfer with subsequent removal of the selection gene from the host genome. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10558–10562. doi: 10.1073/pnas.88.23.10558. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dale E. C., Ow D. W. Intra- and intermolecular site-specific recombination in plant cells mediated by bacteriophage P1 recombinase. Gene. 1990 Jul 2;91(1):79–85. doi: 10.1016/0378-1119(90)90165-n. [DOI] [PubMed] [Google Scholar]
- Golic K. G. Site-specific recombination between homologous chromosomes in Drosophila. Science. 1991 May 17;252(5008):958–961. doi: 10.1126/science.2035025. [DOI] [PubMed] [Google Scholar]
- Henikoff S. Position effect and related phenomena. Curr Opin Genet Dev. 1992 Dec;2(6):907–912. doi: 10.1016/s0959-437x(05)80114-5. [DOI] [PubMed] [Google Scholar]
- Herman R. K. Crossover suppressors and balanced recessive lethals in Caenorhabditis elegans. Genetics. 1978 Jan;88(1):49–65. doi: 10.1093/genetics/88.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Imai T., Olson M. V. Second-generation approach to the construction of yeast artificial-chromosome libraries. Genomics. 1990 Oct;8(2):297–303. doi: 10.1016/0888-7543(90)90285-3. [DOI] [PubMed] [Google Scholar]
- Lichtenstein C., Barrena E. Prospects for reverse genetics in plants using recombination. Plant Mol Biol. 1993 Mar;21(6):v–xii. doi: 10.1007/BF00023619. [DOI] [PubMed] [Google Scholar]
- Matsuzaki H., Nakajima R., Nishiyama J., Araki H., Oshima Y. Chromosome engineering in Saccharomyces cerevisiae by using a site-specific recombination system of a yeast plasmid. J Bacteriol. 1990 Feb;172(2):610–618. doi: 10.1128/jb.172.2.610-618.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Odell J., Caimi P., Sauer B., Russell S. Site-directed recombination in the genome of transgenic tobacco. Mol Gen Genet. 1990 Sep;223(3):369–378. doi: 10.1007/BF00264442. [DOI] [PubMed] [Google Scholar]
- Orban P. C., Chui D., Marth J. D. Tissue- and site-specific DNA recombination in transgenic mice. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6861–6865. doi: 10.1073/pnas.89.15.6861. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Russell S. H., Hoopes J. L., Odell J. T. Directed excision of a transgene from the plant genome. Mol Gen Genet. 1992 Jul;234(1):49–59. doi: 10.1007/BF00272344. [DOI] [PubMed] [Google Scholar]
- Saghai-Maroof M. A., Soliman K. M., Jorgensen R. A., Allard R. W. Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci U S A. 1984 Dec;81(24):8014–8018. doi: 10.1073/pnas.81.24.8014. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sauer B., Henderson N. Cre-stimulated recombination at loxP-containing DNA sequences placed into the mammalian genome. Nucleic Acids Res. 1989 Jan 11;17(1):147–161. doi: 10.1093/nar/17.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sauer B., Henderson N. Site-specific DNA recombination in mammalian cells by the Cre recombinase of bacteriophage P1. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5166–5170. doi: 10.1073/pnas.85.14.5166. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sauer B. Identification of cryptic lox sites in the yeast genome by selection for Cre-mediated chromosome translocations that confer multiple drug resistance. J Mol Biol. 1992 Feb 20;223(4):911–928. doi: 10.1016/0022-2836(92)90252-f. [DOI] [PubMed] [Google Scholar]
- van Haaren M. J., Ow D. W. Prospects of applying a combination of DNA transposition and site-specific recombination in plants: a strategy for gene identification and cloning. Plant Mol Biol. 1993 Nov;23(3):525–533. doi: 10.1007/BF00019300. [DOI] [PubMed] [Google Scholar]