DNA

Part:BBa_K4610007:Design

Designed by: Sophie Geierstanger   Group: iGEM23_CCA-San-Diego   (2023-10-08)


Afu8g00470 (Fma-ABM)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 310
    Illegal PstI site found at 197
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 310
    Illegal PstI site found at 197
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 310
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 310
    Illegal PstI site found at 197
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 310
    Illegal PstI site found at 197
    Illegal AgeI site found at 480
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

In order to increase the rate of transfection, our team has decided to construct two 4 layer chitosan nanocomposites that contain in total 8 DNA sequences rather than one 8 DNA layer nanocomposite. The first chitosan nanocomposite, chitosan nanocomposite A, would contain parts BBa_K4610000, BBa_K4610001, BBa_K4610002, and BBa_K4610003 (in order from outer to inner layer). The second chitosan nanocomposite, chitosan nanocomposite B, would contain BBa_K4610004, BBa_K4610005, BBa_K4610006, and BBa_K4610007 (in order from outer to inner layer).

In the constructed chitosan nanocomposite B, BBa_K4610007 is the 4th layer and innermost core layer of the nanoparticle. It is the last DNA segment to be released after the subsequent degradation and release of all previous DNA layers. The successful transfection and transport of the chitosan nanoparticles pDNA layers to the cell's nucleus will result in the production of the DNAs corresponding proteins and ultimately the synthesis of fumagillin.

Source

The source of Afu8g00470 is aspergillus fumigatus, a saprophytic filamentous fungus. Afu8g00470 is located on chromosome 8. Af293 is a clinically derived strain that was isolated in 1994 from a lung biopsy taken post autopsy from a patient who was developing severe sepsis syndrome. In 1998 it became the first A. fumigatus isolate with a full genome sequence and even in 202, Af293 genome is the only genome sequence of A. fumigatus to have a full physical reconstruction of its chromosomes. Af293 has eight chromosomes, however, Afu8g00470 exists on the 8th chromosome.


References

Guruceaga, X., Perez-Cuesta, U., Abad-Diaz de Cerio, A., Gonzalez, O., Alonso, R. M., Hernando, F. L., Ramirez-Garcia, A., & Rementeria, A. (2019, December 20). Fumagillin, a mycotoxin of aspergillus fumigatus: Biosynthesis, biological activities, detection, and applications. Toxins. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7020470/

Bertuzzi, M., van Rhijn, N., Krappmann, S., Bowyer, P., Bromley, M. J., & Bignell, E. M. (2020, September 17). On the lineage of aspergillus fumigatus isolates in common laboratory use. OUP Academic. https://academic.oup.com/mmy/article/59/1/7/5908083