Cloning of Chikungunya Virus E2 Gene in Escherichia coli TOP 10

摘要

Chikungunya virus (CHIKV) infection generally occurs in tropical and temperate regions such as Indonesia and causes a large amount of socio-economic loss. People infected with CHIKV experience severe arthralgia that can last for months to years. Nowadays, there are no low cost and powerful diagnostics system that can effectively prevent and diagnose CHIKV infection. For that reason, this study focuses on development of simple, robust, and rapid diagnostic assay for chikungunya virus infection. The aim of this study is creating recombinant plasmid vector with molecule size 7.223 bp. Here, the 1.260 bp of gene insert envelope 2 (E2) protein CHIKV was cloned in 5.963 bp pYES2/CT vector. This recombinant vector transformed to Escherichia coli TOP 10 then plated in ampicillin selective agar medium. Several positive colonies of E. coli recombinant were isolated to obtain its plasmid molecule. Further step of confirmation recombinant vector analysis was done by digestion and PCR methods. The results of digestion method showed that two DNA bands appear with each size 5.963 bp and 1.260 bp while PCR method showed DNA bands by the size 1.260 bp and 1.560 bp. The conclusion is we have successfully cloned the CHIKV E2 protein in vector plasmid pYES2/CT based on digestion and PCR method.