You need to express and purify a large amount of a protein for testing in an ani

Question

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 “if A doesn’t work I will try B, and I think B is the next best choice because…”.

Explanation / Answer

The protein of interest is going to have a gene coding for it, we should use recombinant DNA technology to insert the target gene in a high copy number plasmid and transform a bacterial cell to express the protein. The purification of the protein can be done by chromatography. If the expression of the protein is not high we can change the vector like use pBR322 that is traditionally used or try with a more copy number plasmid pUC19 500-700 copies.

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