Expression, Isolation, and Characterization of Tag-less Human Interleukin 6 in Salt inducible Bacterial System; GJ-1158
Keywords:
human interleukin-6, Salt Induction, GJ1158 Expression Host, IL-6- a Biomarker, Characterization of hIL-6 and Target for various disorders.Abstract
Background: hIL-6 is a cytokine that is produced in response to inflammation and tissue injury. It has pleiotropic effect and is composed of 212 amino acids. Recently, in COVID-19, breast cancer, gastric cancer etc hIL-6 became a target for chemotherapy. It is also used as a biomarker for various autoimmune and viral diseases. It is already expressed in, many systems but E.coli, GJ1158 is suitable for the expression of hIL-6. Furthermore, we performed an in silico docking study in the presence of plant-derived anti-inflammatory alkaloid Mandindoline A to probe the potential hIL-6 interaction. This can have implications for the next anti-IL-6 treatments.
Objective: Here in this work the hIL-6 gene was subcloned into pRSET-B from the mammalian expression vector pCDNA 3.1 and expressed in a salt inducible expression system GJ1158 with help of sodium chloride instead of IPTG as an inducer.
Methodology: Protein is getting expressed in soluble as well as aggregate form. The soluble fraction was obtained through freeze-thaw cycles and subsequently lyophilized. Those soluble parts of protein get loaded on the 12% SDS-gel. The lyophilized sample used for the purification and MALDI characterization.
Results: Human interleukin-6 was expressed on a 12% SDS gel at nearly 23.7 kDa and get confirmed by Western blotting by using anti-interleukin-6 antibody.
Conclusion: hIL-6 is successfully produced from a salt inducible system GJ1158. As per best of our knowledge, till now there is no record found for the expression of tagless hIL-6 in GJ1158.
