The Gin recombinase of phage Mu can catalyse site-specific recombination in plant protoplasts.
A mutant Gin recombinase of the phage Mu DNA inversion system was successfully expressed in Arabidopsis thaliana and tobacco protoplasts. Site-specific recombination was monitored both physically and biologically with the help of a recombination assay system in which expression of a beta-glucuronidase (gus) gene requires Gin-mediated recombination. We demonstrate that the wild-type Gin protein is not able to promote recombination in plant protoplasts, presumably because plant cells do not contain a protein that can substitute for the Escherichia coli FIS protein needed for full activity of wild-type Gin in E. coli. A FIS-independent Gin mutant protein on the other hand was efficient in promoting recombination on recombination substrates introduced transiently and on substrates stably integrated into the plant genome. We discuss the various advantages this system can provide for genetic manipulation of plant cells.
Maeser
S
S
Institut für Genbiologische Forschung Berlin GmbH, FRG.
Kahmann
R
R
D010583:Bacteriophage mu / Q000201:enzymology*; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D015870:Gene Expression; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D010957:Plasmids; D011523:Protoplasts / Q000378:metabolism*; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic
D004247:DNA; D004254:DNA Nucleotidyltransferases; C071957:Gin recombinase