Abstract
Dengue is an infectious disease affecting 390 millions people in tropical and subtropical region annually. Dengue virus NS1 protein plays an important role in viral replication in the host cell and it is detected in high level in the infected patient serum. A synthetic gene of untagged DENV-2 NS1 with codons optimization for expression in Escherichia coli has been generated and then inserted into an expression vector pET16b. As a hydrophobic protein with aliphatic index of 71.21%, DENV-2 NS1 was produced as inclusion bodies in E. coli BL21(DE3). The DENV-2 NS1 aggregate was unfolded in 8 M urea and then refolded by reverse dilution method. The refolded DENV-2 NS1 has immunologically active structure as it is capable to interact with anti-NS1 antibody, hence making it as a potential vaccine candidate.
Highlights
Disease caused by the Dengue Virus (DENV) varies from asymptomatic, non-specific fever, Dengue Fever (DF) to a deadly form of Dengue Hemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS) (Guzman et al, 2013)
We reported the expression of untagged DENV-2 NS1 in E. coli BL21(DE3)
DENV-2 NS1 gene was synthesized with codon optimization for E. coli in order to achieve the best protein expression
Summary
Disease caused by the Dengue Virus (DENV) varies from asymptomatic, non-specific fever, Dengue Fever (DF) to a deadly form of Dengue Hemorrhagic Fever (DHF) and Dengue Shock Syndrome (DSS) (Guzman et al, 2013). The seven non-structural proteins of DENV are NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5 (Muller and Young, 2013). The non-structural protein NS1, plays an important role in the human immune invasion and evasion mechanisms (Rastogi et al, 2016), RNA replication in the host cell (Sampath and Padmanabhan, 2009; Fan et al, 2014), production of infectious viral particles (Scaturo et al, 2015), autoimmune damage of the host tissue (Amorim et al, 2014) and is a potential protein for vaccine candidate Most attempts to produce recombinant NS1 used a tag or fusion system, such as His tag or thioredoxin, which requires removal of the tag or fusion partner, especially when the NS1 is designated for a vaccine candidate In this studies, we reported the expression of untagged DENV-2 NS1 in E. coli BL21(DE3). The refolding process to obtain an immunological active DENV-2 NS1 recognized by NS1 antibody is described
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