Abstract
The Cre-lox site-specific recombination system of bacteriophage P1 was used to excise a firefly luciferase (luc) gene which had previously been incorporated into the tobacco genome. The excision event was due to site-specific DNA recombination between two lox sequences flanking the luc gene and was catalyzed by the Cre recombinase introduced by cross-fertilization. Recombination resulted in the fusion of a promoter with a distally located hygromycin phosphotransferase (hpt) coding sequence and the excision event was monitored as a phenotypic change from expression of luc to expression of hpt. The efficiency of recombination was estimated from the exchange of gene activity and confirmed by molecular analysis. The relevance to potential applications of site-specific deletion-fusion events for chromosome engineering are discussed.
Publication types
- Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
- Agrobacterium tumefaciens / enzymology
- Agrobacterium tumefaciens / genetics*
- Bacteriophages / enzymology
- Bacteriophages / genetics
- Base Sequence
- Blotting, Northern
- Cinnamates*
- DNA Nucleotidyltransferases / genetics*
- DNA Nucleotidyltransferases / metabolism
- Genetic Engineering / methods
- Hygromycin B / analogs & derivatives
- Hygromycin B / pharmacology
- Integrases*
- Luciferases / genetics
- Luciferases / metabolism
- Molecular Sequence Data
- Nicotiana / drug effects
- Nicotiana / enzymology
- Nicotiana / genetics*
- Phosphotransferases (Alcohol Group Acceptor)*
- Phosphotransferases / genetics
- Phosphotransferases / metabolism
- Plants, Genetically Modified / drug effects
- Plants, Genetically Modified / enzymology
- Plants, Genetically Modified / genetics*
- Plants, Toxic*
- Polymerase Chain Reaction
- Recombination, Genetic / genetics*
- Viral Proteins*
Substances
- Cinnamates
- Viral Proteins
- Hygromycin B
- hygromycin A
- Luciferases
- Phosphotransferases
- Phosphotransferases (Alcohol Group Acceptor)
- hygromycin-B kinase
- Cre recombinase
- DNA Nucleotidyltransferases
- Integrases