Abstract
A dengue virus serotype 3 strain (DENV-3) was isolated in Jakarta, Indonesia. The NS3 subunit of this strain was selected for recombinant expression to determine its suitability toward later developing a subunit recombinant vaccine for dengue hemorrhagic fever. NS3 is a non-structural protein weighing approximately 72 kDa. It plays a key role in the replication cycle of the dengue virus. The successful cloning of the DENV-3 NS3 coding sequence into a pYES2/CT vector was confirmed, and Saccharomyces cerevisiae cells were transformed with this vector, followed by the expression and purification of the recombinant protein. PCR amplification and DNA sequencing confirmed the presence of the NS3 gene, and SDS-PAGE and western blot analysis produced a specific band of ∼72 kDa.
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