But also, for those with modest experience in the production of heterologous proteins, we describe the many options and approaches that have been successful for expressing a great number of proteins over the last couple of decades, by answering the questions needed to be addressed at the beginning of the project. For this reason, in this review, we comment on the most recent advances in the topic. Yet, in the field of recombinant protein expression and purification, progress is continuously being made. Collectively, these papers gather more than 2000 citations. In the past, many reviews have covered this topic with great detail ( Makrides, 1996 Baneyx, 1999 Stevens, 2000 Jana and Deb, 2005 Sorensen and Mortensen, 2005). Poor growth of the host, inclusion body (IB) formation, protein inactivity, and even not obtaining any protein at all are some of the problems often found down the pipeline. In practice, however, dozens of things can go wrong. You take your gene of interest, clone it in whatever expression vector you have at your disposal, transform it into the host of choice, induce and then, the protein is ready for purification and characterization.
The ability to express and purify the desired recombinant protein in a large quantity allows for its biochemical characterization, its use in industrial processes and the development of commercial goods.Īt the theoretical level, the steps needed for obtaining a recombinant protein are pretty straightforward.
FINALE 2014.5 CHANGING DIFFICULTY LEVEL FOR INSTRUMENT HOW TO
Every researcher that embarks on a new project that will need a purified protein immediately thinks of how to obtain it in a recombinant form. The days where kilograms of animal and plant tissues or large volumes of biological fluids were needed for the purification of small amounts of a given protein are almost gone. There is no doubt that the production of recombinant proteins in microbial systems has revolutionized biochemistry.
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