Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of dengue 2 virus in Pichia pastoris | Zendy

Fernita Puspasari | Zendy; Riski Dwimalida Putri | Zendy; Aisyah Aisyah | Zendy; Raden Roro Rika Damayanti | Zendy; Anita Yuwita | Zendy; Bachti Alisjahbana | Zendy; Sukwan Handali | Zendy; Ihsanawati Ihsanawati | Zendy; Dessy Natalia | Zendy

Open Access

Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of dengue 2 virus in Pichia pastoris

Author(s) -

Fernita Puspasari,

Riski Dwimalida Putri,

Aisyah Aisyah,

Raden Roro Rika Damayanti,

Anita Yuwita,

Bachti Alisjahbana,

Sukwan Handali,

Ihsanawati Ihsanawati,

Dessy Natalia

Publication year - 2017

Publication title -

asian pacific journal of tropical biomedicine

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.507

H-Index - 61

eISSN - 2588-9222

pISSN - 2221-1691

DOI - 10.1016/j.apjtb.2017.07.017

Subject(s) - pichia pastoris , recombinant dna , dengue virus , antigenicity , pichia , microbiology and biotechnology , biology , virology , gene , expression vector , dengue fever , antibody , biochemistry , genetics

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

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