Briefly describe the objective of the experiment outlined in the paper. Briefly explain how a protein with a polydistidine tag is created. What metal is used to purify dioxygenase? Compare the wash buffer and the elution buffers. What strategy is being used to elute the protein from the affinity column? Explain. The Creation of a Protein with a Polydistidine Tag

A Polydistidine Tag is used to create a protein

Proteins are vital building blocks of life. They are also responsible for many important body functions. A tag is a common way to identify proteins and make it easier to manipulate them. The most common tag for protein research is the polydistidine tag. There are several steps involved in creating a polydistidine-tagged protein. The first step is to insert the gene that encodes the desired protein into an expression vector. This vector is then translated into host cells. A polydistidine tag is then used to express the desired protein in the host cell. A polydistidine tag typically contains three to five residues of histidine that link to the protein through a peptide bonds. The host cell is used to purify the tagged protein using affinity chromatography and two-dimensional gel electrophoresis. The tagged protein is then purified from the host cell using affinity chromatography or two-dimensional gel electrophoresis.
Because of their simplicity and stability, polydistidine tags are becoming more popular in the last few years. Studies have shown that polydistidine tags exhibit high levels of stability over a wide range of temperatures (Vieira et al., 2016; Castillo-González et al., 2021). Additionally, these tags allow for rapid, efficient and effective purification of complex proteins (Matsunaga-Gonzalez et. al., 2019). Cont….