Molecular cloning is a process that manipulates the spider silk sequence to select for a proper sequence size and specific vector into which it can be expressed. This process is achieved using techniques such as digests, transformations, purifications, and ligations. This semester, our work has centered around the mPRI(alfalfa), pOET2(insect), and pmk(ecoli) vectors, into which we have, or will, insert 3 time, 6 time, and 9 time sequences of the spider silk amino acid chain. With each repetition of the silk sequence, its properties improve and become more like natural silk. Vector specification allows these sequences to be produced in different hosts, with each providing beneficial and detrimental circumstances to the silks eventual development and production. These processes are ongoing but we hope to produce usable protein in both the mPRI and pOET2 vectors in the coming months, allowing us to refine our production process until the best possible product is achieved.
Worley, Josh, "Molecular Cloning" (2017). Biology Posters. Paper 198.