In Silico Design and Expression of Dengue Virus Type 2 Envelope Domain III in Escherichia coli BL21 (DE3) for Specific Diagnostics
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Date
2026-06-30
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Publisher
MUST Journal of Research and Development (MJRD)
Abstract
Dengue virus type 2 (DENV-2) is a mosquito-borne virus of global
public health concern. Routine diagnostic tests often target whole
envelope glycoproteins that suffer from cross-reactivity. The
envelope domain III (EDIII) protein contains subtype virus-specific
epitopes, offering improved specificity over complete envelope.
This study evaluated a modified DENV-2 EDIII expression in E. coli
BL21 (DE3) using a codon-optimized construct. AlphaFold2
algorithms were utilized to predict its 3D structure and identify
potential antigenic epitopes. The construct was stable (instability
index <40), soluble and antigenic (>0.5score, respectively) containi
ng three key linear epitopes; CTGKFKVVKEIAETQH (302–317aa),
SPCKIPFEIMDLEKR (331–345aa), and QLKLNWFKKGSS (386–
396aa). Optimal expression occurred at 30°C and 4 hours post-
induction, producing a distinct 14 kDa band on SDS-PAGE
consistent with the expected molecular weight of target EDIII
protein. Overall, these findings demonstrate that in silico protein
design with low-temperature bacterial expression can produce
recombinant proteins suitable for specific diagnostics
Description
Keywords
n silico design, Recombinant expression, EDIII, DENV-2