title: Use of the Cre-loxP recombination system as an estimate for Agrobacterium-mediated co-transformation of tobacco leaves.
abstract: Agrobacterium tumefaciens-mediated transformation of tobacco leaves (Nicotiana tabacum) is used to study gene expression in a heterologous genetic background. Here, the Cre-loxP recombination system was used to detect T-DNA transfer by two A. tumefaciens cells harboring different binary vectors. Cre, under the control of the CaMV 35S promoter, was cloned into one vector, and a loxP cassette into another vector. A mixture of A. tumefaciens, in which each cell contained either a Cre- or loxP-vector, was co-infiltrated into tobacco leaves. After two days, excision of loxP-flanked DNA was detected by PCR and used as an estimate for co-transformation events. Strongest excision (> 50%) was observed when the loxP cassette was cloned into vector pPZP112 and Cre into pISV2678. This fast and easy technique can be used to assess the co-transformation efficiency of tobacco cells in future studies.
authors: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen (Zhongshan) University, East Campus, Guangzhou, 510006, China.
authors: Min-Ting
authors: MT
authors: Liang
authors: Cui-Ping
authors: CP
authors: Yang
authors: Zhi-Ping
authors: ZP
authors: Xie
authors: Christian
authors: C
authors: Staehelin
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D017796:Caulimovirus; D004269:DNA, Bacterial / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D008967:Molecular Biology / Q000379:methods; D018515:Plant Leaves / Q000235:genetics / Q000382:microbiology; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D014026:Nicotiana / Q000235:genetics* / Q000382:microbiology*; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA