Difference between revisions of "Part:BBa K678063"

Latest revision as of 19:00, 27 September 2011

pJEJAM15

pJEJAM15 is a plasmid that after digestion with NotI is intended to be transformed into Aspergillus nidulans and integrated into the genome by non-homologous end joining. The plasmid consists the strong constitutive gpdA promoter , mRFP with a nucleosomal targeting sequence (NLS) fused to the C-terminus, the trpC terminator, a pyrG marker cassette for selection, an ori for propagation in E. coli, and an ampR resistance cassette (Figure 1).

DTU-Denmark-2 2011 Figure 1: Plasmid map of pJEJAM15, the figure is not drawn to scale.

Red fluorescence can be observed in clear spots in Figure 3. When comparing Figure 2 and FIgure 3 it can be seem that both spores and hyphae fluoresce. We assume that the fusion of mRFP1 with the NLS results in the targeting to the nucleus. To confirm that the mRFP1 in fact is targeted to the nucleus further experiments would have to be conducted. In figure 4 and figure 5 it be can see that the integration of the linearized pJEJAM15 must have impaired a gene or pathway of the transformant because its morphology and growth differed slightly from the [http://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild%20type wild-type].

DTU-Denmark-2 2011 Figure 2: Differential interference contrast image of Aspergillus nidulans with pJEJAM15.

DTU-Denmark-2 2011 Figure 2: Fluorescence image of Aspergillus nidulans pJEJAM15.

DTU-Denmark-2 2011 Figure 2: Aspergillus nidulans Front

DTU-Denmark-2 2011 Figure 2: Aspergillus nidulans Back

Sequence and Features