Question

You are interested in whether the protein you are studying for your dissertation, ZyxR, is capable of binding to and regulating expression of the ZyxABC operon. ZyxR is a 24kDa protein with a slightly negative overall charge and no known inherent tags. ZyxR is known to bind to the zyxY gene to regulate expression. yopA (100bp) is not regulated by ZyxR. The first step in this process is to determine whether ZyxR is capable of binding to the regulatory regions surrounding the zyxABC operon.

Give one potential strategy that you could use to purify ZyxR. Your strategy should be a multistep process beginning with cell lysate (containing millions of proteins) that you will take to purify the protein, should address potential shortcomings of each step, and should attempt to rectify those shortcomings. If your strategy requires any special conditions or changes to ZyxR, you should include those in the details. Drawings of your strategy/steps are highly encouraged!

Answer 1

Step-by-step explanation:

One potential methodology which could be utilized to cleanse ZxyR is Affinity chromatography. Affinity chromatography is utilized for the most part relevant methods for cleansing proteins. This strategy exploits the high harmony of numerous proteins for explicit synthetic gatherings. For instance, the plant protein concanavalin A can be filtered and purified by going a rough concentrate crude extract through a segment of beads encapsulating covalently joined glucose deposits.