Cloning and Expression of Non-structural I (NSI) Protein of Zika Virus

Low, Yong Chun (2018) Cloning and Expression of Non-structural I (NSI) Protein of Zika Virus. [Final Year Project Report] (Unpublished)

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Abstract

Zika virus (ZIKV) belongs to the family Flaviviridae. It is spread through the bite of an infected mosquito, especially of the Aedes aegypti. The transmission of ZIKV to the foetus may cause birth defect in some cases. The purpose of conducting this study is to clone and express the nonstructural l (NS 1) protein of the ZIKV and purify the protein for antigenicity study. NS 1 protein is involve in the infectious viral production and pathogenesis of the Zika disease. The gene encodes the NS 1 protein was amplified using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and cloned in an Escherichia coli (E. coli) cloning vector system, pET SUMO vector system. The plasmid was transformed into the E. coli cloning host, One Shot Mach 1-T1 Competent Cell and subsequently into BL21(DE3) One Shot Competent Cell for protein expression. Once the colony was verified by colony PCR, the expression of the NS 1 recombinant protein was analysed using SDS-PAGE and Western blot. In the study, the NS 1 recombinant protein was successfully expressed and purified. The concentration of the purified recombinant protein was 0.817 mg/ml. Preliminary testing of the NS 1 recombinant protein shows that it has potential to be used for further development of clinical diagnosis and development of specific antibody.

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