This approach also allowed us to examine unintended effects resulting from http://www.selleckchem.com/products/nutlin-3a.html the expression Inhibitors,Modulators,Libraries of Cre recombinase. We found that Cre recombinase had a minimal impact on the transcriptome, which suggests that it produces few unintended effects in Arabidopsis. Microarray analysis of Arabidopsis plants overexpressing ABF3 demonstrated that the impact on the transcriptome is minimal. In the absence of drought stress, there were no differentially expressed genes. In response to drought stress, a reprogramming of the drought response was observed, suggestive of changes in the timing or strength of expression of some genes in 35S ABF3 plants. Some of these changes may be directly related to the action of ABF3 while others may reflect an altered physiological state as a result of the enhanced drought tolerance of Inhibitors,Modulators,Libraries 35S ABF3 plants.

Amongst the differentially expressed genes, no unintended pathways appeared to be activated as a result of ABF3 overexpression. These Inhibitors,Modulators,Libraries results are sig nificant because they demonstrate that plant responses to abiotic stresses such as drought may be strictly coordi nated at multiple regulatory steps and this limits the extent of unintended pleiotropic effects. This demon strates that engineering stress tolerance through manipu lation of endogenous plant pathways may not necessarily produce unintended pleiotropic effects despite the com plexity of such traits. This is an important finding for establishing the safety of such traits as they begin to enter the market in the near future.

Methods Generation of transgenic plants To generate 35S ABF3 plant lines, Arabidopsis thaliana Inhibitors,Modulators,Libraries Col 0 plants Inhibitors,Modulators,Libraries were transformed by the floral dip method using Agrobacterium tumefaciens strain GV3101 harbouring the pCAMBIA3300 vector containing the Cauliflower mosaic virus 35S promoter, ABF3 coding region, and nopaline synthase transcriptional terminator, bordered on either side loxP sites. Homozygous single insert transformants were identified by Southern blot and segregation analysis and three lines with high levels of ABF3 expression, as determined by RT PCR analysis, were selected for further analysis. To generate plant lines expressing Cre recombinase, Arabidopsis thaliana Col 0 plants were transformed as above with a pCAMBIA1200 vector containing the Cau liflower Mosaic Virus 35S promoter, Cre recombinase coding region, and nopaline synthase transcriptional terminator.

Rapamycin mTOR To generate control plant lines, 35S ABF3 plants were crossed with plants expressing Cre recombinase in order to excise the 35S ABF3 nos transgene. The F1 plants were PCR genotyped to identify those that underwent successful excision, using the primers ABF F and Nos R which are specific for the ABF3 gene and the nopaline synthase transcrip tional terminator, respectively. One plant from each cross, Control 48. Cre 1. 1. 2, Control 57. Cre 1. 2.