You need to express and purify a large amount of a protein for testing in an animal trial. The protein is larger than average, though not excessive so (~100 kDa, so the transcript is ~4,000 nucleotides long) and has an unusually acid isoelectric point (pI = 5). Propose and explain a strategy to clone, express, and purify this protein. [note: do not restate your answer for the previous question here, just state what factors you would consider when deciding on a cloning method. There is no single right answer for the choice of expression system or purification strategy; you should however be able to describe a rational approach and state why you chose that approach. Include some discussion of alternatives, such as âif A doesnât work I will try B, and I think B is the next best choice becauseâ¦â.
You need to express and purify a large amount of a protein for testing in an animal trial. The protein is larger than average, though not excessive so (~100 kDa, so the transcript is ~4,000 nucleotides long) and has an unusually acid isoelectric point (pI = 5). Propose and explain a strategy to clone, express, and purify this protein. [note: do not restate your answer for the previous question here, just state what factors you would consider when deciding on a cloning method. There is no single right answer for the choice of expression system or purification strategy; you should however be able to describe a rational approach and state why you chose that approach. Include some discussion of alternatives, such as âif A doesnât work I will try B, and I think B is the next best choice becauseâ¦â.
