Cloning of chikungunya virus envelope 2 (E2) gene to pPICZaA in Escherichia coli TOP10

Abstract

Chikungunya virus infection (CHIKV) causes symptoms of chikungunya fever and joint pain. The chikungunya virus is spread by the bite of an Aedes mosquito. The symptoms of CHIKV and dengue virus (DENV) infection are similar and are spread by the same vector. Diagnosis of CHIKV infection is carried out by expensive molecular detection and immunological detection (RDT) as an alternative diagnosis. The material to be used for the development of RDT CHIKV is the envelope 2 protein (E2) CHIKV. This study aims to obtain pPICZaA-E2 which is transformed into Escherichia coli TOP10. The pPICZaA plasmid and the E2 CHIKV gene were cloned into E. coli TOP10 and grown onto LB+zeocin agar medium. Cultures grown on the medium were verified for colonies carrying pPICZaA-E2 using PCR colony and restriction. The PCR verification results of the colonies from the growing cultures showed a band measuring 1.260 bp. The results of the restriction verification obtained colonies with two bands measuring 3.569 and 1.260 bp. It was concluded that the E. coli TOP10 colonies carried pPICZaA-E2. Sequencing of the isolated pPICZaA-E2 plasmid is required.