Agroinfiltration as a tool for transient expression of cre recombinase in vivo.
Agroinfiltration was used to express transiently cre recombinase from bacteriophage P1 in planta. Activation of gfp expression after cre-mediated excision of a bar intervening sequence served as a marker to monitor site-specific recombination events in lox-target N. benthamiana plants. Gfp expressing regenerants from A. tumefaciens infiltrated leaves were obtained with an efficiency of about 34%. In 20% of the regenerants bar gene excision was due to the expression of stably integrated cre gene, whereas in 14% of plants site-specific recombination was a consequence of transient cre expression. Phenotypic and molecular data indicated that the recombined state has been transferred to the T(1 )generation. These results demonstrate the suitability of agroinfiltration for the expression of cre recombinase in vivo.
Kopertekh
Lilya
L
Federal Biological Research Centre for Agriculture and Forestry, Institute for Plant Virology, Microbiology and Biosafety, Messeweg 11-12, D-38104 Braunschweig, Germany.
Schiemann
Joachim
J
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017101:Bacteriophage P1 / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000235:genetics; D045522:Recombinases / Q000235:genetics; D011995:Recombination, Genetic; D014026:Nicotiana / Q000235:genetics*
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D011994:Recombinant Proteins; D045522:Recombinases; C036483:T-DNA; D049452:Green Fluorescent Proteins