Downstream Processing of Recombinant Proteins in E.coli Essay

Downstream Processing of Recombinant Proteins in E.coli - Essay Example From this study it is clear that hence for the high recovery of the active protein molecule, solubilization and refolding parts must be of high precision. Inclusion bodies consist of polypeptides of the recombinant protein. They are the inactive secondary –like structures. Thus, the isolation and purification of the protein are simple. The activity of the unfolded protein can be brought out by using mild solubilizing conditions. This will help in the high recovery of the bioactive protein than that compared to solubility using high chaotropic agent. The following Downstream processing steps can be used for the production of the recombinant protein from the inclusion bodies The recombinant E.coli is grown in LB medium with antibiotics such as kanamycin or ampicillin or chloramphenicol based on the plasmid. The flasks are shaken at rpm around 150 -250 and the temperature is maintained at 37 degree Celsius. After the cells have reached the log phase, This study outlines that IPTG is added and further shaken for 4-6 hours. The cells are centrifuged and re-suspended in the 50 mM sodium phosphate buffer. The cells are lysed using the homogenizer or sonicator. The cell suspension is centrifuged and filtered using 0.45 Â µm polyethersulfonate membrane. Many protein-specific methods are available for the increased solubility of the recombinant proteins in E.coli. To recover the soluble proteins, strong denaturants like urea, guanidinium hydrochloride are used. The solubilization is carried out under reducing conditions. These inclusion bodies must be washed well before solubilization. Solubilizing agents such as thioredoxin are known to improve the solubility of the proteins. Gene fusion techniques are equally good for the separation of the proteins. Maltose binding protein and Glutathione-S- transferase are found to bind well with the protein and can be removed by using the affinity chromatography techniques. Refolding is performed using dilution or diafliltrat ion in buffers of low.

Finding a scripture Assignment Example | Topics and Well Written Essays - 750 words - 2

Finding a scripture - Assignment Example Nevertheless, the hindrance that wealth has on the walk with Jesus Christ is also confirmed. Salvation is also underscored as impossible for man, but only possible with God. In this pericope, it is clear that God has endowed people with different gifts at varying degrees, and in this regard, everyone is a steward. Rewards are accorded to those who have been faithful with their gifts or talents. From the foregoing, it is clear that money in itself is amoral: it is the love of it that inspires evil. As a corollary to this, money can be used by Christians to extend the Kingdom of God, specifically for the cause of the fulfillment of the Great Commission. God is the Originator of the state and state power and as such, Christians are obligated to observe the laws of the state. Duties which are relevant to the state and sanctioned by the state are concomitant with God’s will. Although God is the Originator of the state and state power (as previously seen in Romans 13:1-7), yet there are instances in which state actors and civil bodies may contravene God’s will, commands or word in their laws or decrees. In this case, the Christian owes his first and foremost allegiance to God. In these situations, God is still sovereign over man’s laws. As already mentioned, in instances in which the state and its actors may contravene God’s will, commands or word in their laws or decrees, the Christian owes his first and foremost allegiance to God. However adverse these situations may be, God is still sovereign over man’s laws. The state is also a creation of God and the Christian is to observe all his civic duties, for God is the author of all authority. However, a Christian’s first and utmost allegiance is to God. Doing the will of God should override that of the state, in the event that the state has made laws that are contrary to God’s will. Even in these situations, God is still sovereign and is not limited to human laws to interact