Expression and Purification of E2 Protein of Chikungunya Virus in Escherichia coli System

• Main Author: Nur Aimi Aizzah, Azmi
• Format: Final Year Project Report
• Language: English
• Published: Universiti Malaysia Sarawak 2017
• Subjects: QR Microbiology
• Online Access: http://ir.unimas.my/id/eprint/30985/2/Nur%20Aimi%20Aizzah.pdf; http://ir.unimas.my/id/eprint/30985/

Chikungunya virus (CHIKV) is a virus that affects human and it is transmitted by mosquitoes. Similar to dengue virus (DENV) infection, the clinical manifestation of the disease includes high fever, chills, rashes, joint and muscle pain. Since dengue infection is endemic in Malaysia, the diagnosis of CHIKV infection is crucial to differentiate DENV and CHIKV infection. To date, the diagnosis of CHIKV infection can only be done at Institute of Medical Research (IMR) and National Public Health Laboratory (NPHL). Both of the laboratories are situated in Peninsular Malaysia and therefore, the diagnosis of CHIKV infection can be delayed. In this study, E2 DNA plasmid was transformed into BL21 (DE3) One Shot Cells. Once the colony was verified by colony PCR, the expression of the recombinant protein was analyzed by SDS-PAGE. Then, the E2 recombinant protein was purified using ProBond Purification System (Novex, CA, USA). E2 recombinant protein was successfully expressed and purified and the concentration of the purified recombinant protein was 0.2405 mg/ml. Preliminary testing of the E2 recombinant protein shows it has a potential to be used in developing a diagnostic assay.