Recombinant expression and purification of the NS3 subunit of a Dengue Virus Type 3 strain isolated in Jakarta

Abinawanto, S. Pambudi, A. Sholiha

Research output: Contribution to journalConference articlepeer-review

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.

Original languageEnglish
Article number012014
JournalIOP Conference Series: Earth and Environmental Science
Volume481
Issue number1
DOIs
Publication statusPublished - 27 Apr 2020
EventLife and Environmental Sciences Academics Forum 2018, LEAF 2018 - Depok, West Java, Indonesia
Duration: 1 Nov 2018 → …

Keywords

  • DENV-3
  • NS3 protein
  • plasmid pYES2/CT
  • Saccharomyces cerevisiae

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