Open AccessConstruction 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,
Raden Roro Rika Damayanti,
Anita Yuwita,
Bachti Alisjahbana,
Sukwan Handali,
Ihsanawati,
Dessy Natalia
Publication year - 2017
Publication title -
asian pacific journal of tropical biomedicine/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 , dengue fever , dengue virus , gene , virology , glycoprotein , dengue hemorrhagic fever , gene expression , biology , virus , pichia , computational biology , genetics , recombinant dna
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