Difference between revisions of "Part:BBa K678060"

Latest revision as of 15:42, 27 September 2011

pJEJAM12

pJEJAM12 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 (NHEJ). The plasmid consists the strong constitutive gpdA promoter, green fluorescent protein (GFP), 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 pJEJAM12, the figure is not drawn to scale.

When pJEJAM12 was integrated into A. nidulans expression of the gene encoding GFP, resulted in an even distribution of GFP in the hyphae (Figure 3). When comparing figure 2 and figure 3 it can be seem that GFP was not expressed in all hyphae, this could be as a result of the integration by NHEJ as we in figure 3 and figure 4 can see that the integration must have impaired a gene or pathway of the transformant, a picture of the wild-type strain can be found [http://2011.igem.org/Team:DTU-Denmark-2/results/Proofofconcept/fungi#Wild%20type here].

DTU-Denmark-2 2011 Figure 2: DIC image of Aspergillus nidulans with pJEJAM12.

DTU-Denmark-2 2011 Figure 3: Fluorescence image of Aspergillus nidulans with pJEJAM12.

DTU-Denmark-2 2011 Figure 4: Aspergillus nidulans Front

DTU-Denmark-2 2011 Figure 5: Aspergillus nidulans Back

Sequence and Features