Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of dengue 2 virus in Pichia pastoris
10.1016/j.apjtb.2017.07.017
- Author:
Fernita PUSPASARI
1
;
Riski Dwimalida PUTRI
1
;
Riski Dwimalida AISYAH
1
;
Raden Roro Rika DAMAYANTI
1
;
Raden Roro Rika IHSANAWATI
1
;
Dessy NATALIA
1
;
Anita YUWITA
2
;
Sukwan HANDALI
2
;
Bachti ALISJAHBANA
3
Author Information
1. Biochemistry Research Division, Faculty of Mathematics and Natural Sciences, Institute of Technology Bandung
2. Pakar Biomedika Indonesia
3. Faculty of Medicine, Universitas Padjadjaran
- Publication Type:Journal Article
- Keywords:
Dengue virus;
DENV 2;
Diagnostic kit;
NS1 protein;
Pichia pastoris
- From:Asian Pacific Journal of Tropical Biomedicine
2017;7(8):689-693
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To express and characterize NS1 of Indonesian-specific DENV2 virus in Pichia pastoris (P. pastoris). Methods A codon optimized synthetic gene derived from the DENV-2 NS1 amino acid sequences was synthesized commercially and inserted into the P. pastoris pPICZαA expression vector. The recombinant DENV-2 NS1 protein was purified by Ni-NTA affinity chromatography, and its antigenicity was tested. Results The recombinant DENV-2 NS1 protein was secreted as a protein with a molecular weight of ∼45 kDa, and the optimal expression condition was achieved by induction with 2% (v/v) methanol for 72 h. The purified recombinant DENV-2 NS1 protein was able to interact with a monoclonal antibody of NS1 in a commercial rapid test. Conclusions The resulting recombinant DENV-2 NS1 protein produced in P. pastoris KM71 is a potential candidate for use in the development of a dengue diagnostic kit and vaccine.
