Seerat Aujla, Angham Ahmed, Dr. Cory Brooks
Volume 7, Issue 1 2023
Page: 20-24
TO introduce and express a protein of interest in the selected host, cloning, transforming, and purifying procedures are performed. Cloning allows production of identical copies of the gene and insert it into a plasmid vector called pET28a(+). This protects the protein’s genetic material from the host’s degradation mechanism. Next, the vector and protein are transformed into a bacterial cell line, or expression host, to propagate and produce the target protein, Siglec-9. Siglec-9 was transformed into BL21(DE3) and Shuffle T7 Express cell lines for expression purposes. After liters of bacterial culture were grown, it underwent a purification process. The goal is to isolate the protein of interest and test for expression with protein electrophoresis and further analysis if needed. The objective is to clone, transform, and purify Siglec-9 for further experimentation.
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