A new technique for activation tagging in Arabidopsis.
We have created and applied to Arabidopsis thaliana a new system of two vectors. The first vector (pEnLox) is intended for activation tagging and contains a multimerized transcriptional enhancer from the cauliflower mosaic virus (CaMV) 35S gene in T-DNA flanked by two loxP-sites and the second vector (pCre) contains the cre gene. Using pEnLox we have generated more than a hundred mutants resistant to the herbicide ammonium glufosinate, and about ten helper-lines resistant to the antibiotic hygromycin obtained with the use of pCre vector and also more than ten double mutants resistant to both selective markers. In at least 3 cases among 40 mutant lines that have been analyzed we observed constitutive ectopic expression of the genes adjacent to the T-DNA insertion that causes development of the mutant phenotype. Also, reversion of the mutants to the wild-type phenotype after removing the CaMV enhancer has been demonstrated. The system presented here provides a new and easier way to analyze A. thaliana gain-of-function mutants.
Pogorelko
Gennady V
GV
NI Vavilov Institute of General Genetics RAS, Moscow, Russia. gpogorelko@yandex.ru
Fursova
Oksana V
OV
Ogarkova
Olga A
OA
Tarasov
Valentin A
VA
D017360:Arabidopsis / Q000235:genetics*; D017796:Caulimovirus / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005822:Genetic Vectors; D019426:Integrases / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA; C045073:Cre recombinase; D019426:Integrases