RESUMO
Using an estrogen-inducible expression XVE (LexA-VP16-Estragon Receptor) system, we have generated approximately 40 000 independent T-DNA insertion lines of Arabidopsis thaliana. Segregation analyses of about 18000 lines indicated that 51.6% of them contain single T-DNA insertions and that the average insertion number is 1.38 copies per line. Mutants displaying a variety of morphological alterations were identified, including those that affect development of roots,hypocotyls, leaves, floral organs and seeds as well as the flowering time.
Assuntos
Arabidopsis/genética , DNA Bacteriano/genética , Mutagênese Insercional/métodos , Plantas Geneticamente Modificadas/genética , Arabidopsis/anatomia & histologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Bactérias/genética , Clonagem Molecular , Estrogênios/farmacologia , Vetores Genéticos/genética , Proteína Vmw65 do Vírus do Herpes Simples/genética , Mutagênese Insercional/efeitos dos fármacos , Fenótipo , Plantas Geneticamente Modificadas/anatomia & histologia , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plasmídeos/genética , Receptores de Estrogênio/genética , Serina Endopeptidases/genética , Ativação Transcricional/efeitos dos fármacosRESUMO
Here, we report our effort in generating an ORFeome collection for the Arabidopsis transcription factor (TF) genes. In total, ORFeome clones representing 1,282 Arabidopsis TF genes have been obtained in the Gateway high throughput cloning pENTR vector, including 411 genes whose annotation lack cDNA support. All the ORFeome inserts have also been mobilized into a yeast expression destination vector, with an estimated 85% rate of expressing the respective proteins. Sequence analysis of these clones revealed that 34 of them did not match with either the reported cDNAs or current predicted open-reading-frame sequences. Among those, novel alternative splicing of TF gene transcripts is responsible for the observed differences in at least five genes. However, those alternative splicing events do not appear to be differentially regulated among distinct Arabidopsis tissues examined. Lastly, expression of those TF genes in 17 distinct Arabidopsis organ types and the cultured cells was profiled using a 70-mer oligo microarray.