Neurotensin receptor 1 (NT1) is over-expressed in up to 35% of all human ductal breast cancers1–2. NT1 provides a target for breast cancer treatment where Neurotensin protein fusions can be designed for diagnostic, marking cancerous cells, and therapeutic responses, treating cancer.
The project focused on producing recombinant DNA for recombinant neurotensin protein fusions. The work involved designing custom primers for the inserted gene and iterations using polymerase chain reaction (PCR). The completed gene DNA sequence coded for Neurotensin and green fluorescence proteins (GFP) where additional iterations were required to add a polyhistidine-tag, HIS-tag, for Nickel column recombinant protein purification.
The project involved electrophoresis and DNA extraction of relevant DNA strands for building the inserted gene. Additionally, the project had transfection, cloning and plating of competent Escherichia coli (E. Coli) cells requiring aseptic techniques.
This was a final year undergraduate project at the University of Lincoln. The supervisor was Dr. Alan Goddard.
References
- Dupouy, S., Viardot-Foucault, V., Alifano, M., Souazé, F., Plu-Bureau, G., Chaouat, M., Lavaur, A., Hugol, D., Gespach, C., Gompel, A., Forgez, P. (2009). The neurotensin receptor-1 pathway contributes to human ductal breast cancer progression. PLoS ONE, 4(1), e4223. https://doi.org/10.1371/journal.pone.0004223
- Souazé, F., Dupouy, S., Viardot-Foucault, V., Bruyneel, E., Attoub, S., Gespach, C., Gompel, A., & Forgez, P. (2006). Expression of neurotensin and NT1 receptor in human breast cancer: A potential role in tumor progression. Cancer Research, 66(12), 6243–6249. https://doi.org/10.1158/0008-5472.CAN-06-0450
