Abstract

The cloned E. coli cell containing Murine IFN -γ inserted pRSET A vector system was effectively expressed in this study. The induction of the clones was done using IPTG in E.coli and induces mRNA generation and synthesis protein. It has shown an expression of protein with 18 kda in SDS PAGE and western blotting and their size was determined by GENE RUNNER software. This recombinant protein has a 6x His tag and it has been proved as it has shown a potent anti His property in western blotting. The purification of the protein was further done by Ni-NTA affinity chromatography. Nitrilo tri acetic acid (NTA) binds more stably with nickel (Ni) with 4 to 6 ligand binding sites in the coordination sphere of Nickel leaving two sites free to interact with the 6X His tag. The total results conclude that the targeted IFN gamma (408bp mouse gene) cloned in pRSET A was effectively expressed in E. coli BL21 strain cells and purified IFN gamma protein effectively as 1mg/ml. The purified IFN gamma protein may be used to diagnose the antiviral activity and antitumor activity. Key words: IFN gamma, pRSET A, E. coli, SDS PAGE, Western Blotting

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.