Cloning and Expression of The Domain III Region of The Envelope Gene of Zika Virus

Easther Mannye, Kennedy (2017) Cloning and Expression of The Domain III Region of The Envelope Gene of Zika Virus. [Final Year Project Report] (Unpublished)

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Abstract

Zika virus (ZIKV) is an arthropod-home virus that causes Zika fever in adults and neurological defects in infants. The severity of the infection ranged from asymptomatic to mild disease and to infection associated with neurological disorders and congenital anomaly. The main objective of this project is to clone the domain III of the envelope gene (E) of ZIKV in E.coli cloning vector. This work is the initial phase of a future study for development of diagnostic assays for detection of ZIKV antibody from human sera. RNA of ZIKV was extracted and envelope gene coding region specifically the Domain III was amplified using reverse-transcription polymerase chain reaction (RT-PCR). The amplified gene was cloned into pET SUMO cloning vector and transformed into Machi-Tl competent E.coli cells. After the selection of the positive transformants that carries the recombinant gene, the envelope gene clones was verified by gene sequencing and transformed into BL21.DE3 competent E.coli for protein expression. The expressed protein was evaluated and analysed by SDS-P AGE and Western blot. At the end of this project, the recombinant protein of expected size 38.2 kDa was obtained.

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