Attributions
Software + Policies & Practices
• PyMOL360 Educational Tool
Patrick Holec
• Synthetic Biology Web Scraper
Patrick Holec
• Mathematical Modeling
Patrick Holec
Wet Lab
Sarah Lucas, Nicholas Nesbitt, Tanner Cook
Wiki
Patrick Holec, Nicholas Nesbitt, Abhiney Jain, Basem Al-Shayeb
Project Support
Basem Al-Shayeb, Sarah Perdue, Casim Sarkar
• Why are we using Pichia pastoris to express Human Insulin?
P. pastoris is a eukaryotic organism in the yeast family that overcomes both obstacles encountered by the aforementioned organisms. P. pastoris has been shown to excrete proteins at a rate of approximately five times that of S. cerevisiae with higher qualitative value as well. Recombinant human insulin has been produced in P. pastoris, but the harvesting and processing procedures are long, complex, and would be greatly simplified through the application of the BioBrick system. P. pastoris is already utilized by the Chinese for various industrial tasks, lending more promise to it as our choice, as industrial application would require very little change to our system.
PCSK1
Figure 1. PCR colony screen of PCSK1-pSB1C3 transformants. Note the banding in each lane at roughly 750 bp, consistent with the second fragment of PCSK1.
Figure 2. PCR colony screen of Ins-pSB1C3 transformants. Banding can be seen at roughly 333 bp, consistent with the full insulin ORF.
Sequencing data received for both PCSK1 and insulin was inconclusive.
Parts List
BBa_K1187001 Human insulin, codon optimized for expression in P. pastoris
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