Part:BBa_K1021007
PtrpC
PtrpC is a strong constitutive promoter from Aspergillus nidulans's tryptophan biosynthesis pathways. It was isolated using a fungal transformation vector and constructed into the pSB1C3 backbone for continuous homologous and heterologous gene expression in fungal chassis.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
To test the activity of the trpC promoter, a composite part was created with GFP downstream (BBa_K1021023). This part was transformed into the ascomycete fungi Cochliobolus heterostrophus. Results indicated that the trpC promoter provided GFP expression within this fungal chassis.
Characterization
DTU-Denmark 2019 characterization The DTU-Denmark 2019 team has characterized the PtrpC promoter in Aspergillus niger. Our characterization experiments were done in two different scales: 1.5 ml wells in a microbioreactor (BioLector), and 10 mL cultures in specialized 50 mL falcon tubes.
GFP expression in Aspergillus niger under PtrpC control
The characterisation of the PtrpC promoter was done by integrating the promoter into the plasmid, as described at the Design page
The promoter was characterised by two separate experiments.
In the first experiment, the PtrpC strain was cultivated in quadruplicates in 50 mL breathable falcon tubes in 10 mL minimal media. A negative control consisting of the parental strain was included. The strains were cultivated for 80 h. at 37C with shaking at 150 rpm. The culture was then stored at 4C, and subsequently protein was extracted and florescence measured as described on the Experiments page. The resulting data can be found below.
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