Nawaf Fawzan Fawzan Alfawzan
Riyadh Schools, Riyadh, SAUDI ARABIA
Due to high efficiency and insolubility, dyes produced by Fluorescent proteins (FP) have been implemented for a variety of applications, such as protein localization and also for studying gene expression. In order to use them, they have to be purified first. However, the purification method being used now, column method, is expensive, time consuming, and has a low efficiency. This project aims to overcome this obstacle by using a new purification method. This was accomplished by a new purification technique that takes advantage of the FPs` high stability against heat; the procedure starts by expressing the protein in a strain of E.coli (BL 21) using the plasmid pET 303. Proceeded by the lysing of the host cell (E.coli). Then the appliance of high amount of temperature to the host cell, resulting in the denaturation of all proteins except for the FPs. Then, an organic solvent (ethanol) was used to remove the organic wastes. As a result, six different fluorescent proteins were purified with a purification percentage ranging between (80% to 100%), with a high yield (max. 480). FPs have a wide range of applications due to their stability under high temperatures, which makes them useful for new studies that aim at using these proteins as catalytic hosts, due to their high stability and organic solvent resistance. In addition, the stability and fluorescence of the FPs will be a target of future enhancement by the mutation of specific amino acids.