Epstein-Barr Virus (EBV) or human herpes virus 4 (HHV-4) is a virus that infects human B cell and leads to nasopharyngeal carcinoma (NPC). The prevention of this disease remains unsuccessful since the vaccine has not been discovered. The objective of this study is to over-produce EBV gp350/220 epitope using several methods in E.coli DH5α. EBV epitope sequences were inserted into pMAL-p5x vector, then transformed into DH5α E.coli and over-produced using 0.3, 1 and 2 mM IPTG. Plasmid transformation was validated using AflIII restriction enzyme in 0.8% agarose. Periplasmic protein was isolated using 2 comparative methods and then analyzed using SDS-PAGE. Method A produced a protein band around 50 kDa and appeared only at transformant. Method B failed to isolate the protein, indicated by no protein band appearing. In addition, any variations in IPTG concentration didn’t give a different result. Thus it can be concluded that even the lowest IPTG concentration is able to induce protein expression.
Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α
Nadya V. M Anyndita, Nurul Dluha, Karimatul Himmah, Muhaimin Rifa’i, Widodo; Comparative method of protein expression and isolation of EBV epitope in E.coli DH5α. AIP Conf. Proc. 29 November 2017; 1908 (1): 060002. https://doi.org/10.1063/1.5012735
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