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

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 re...

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Bibliographic Details
Published in:Asian Pacific Journal of Tropical Biomedicine
Main Authors: Fernita Puspasari, Riski Dwimalida Putri, Aisyah, Raden Roro Rika Damayanti, Anita Yuwita, Bachti Alisjahbana, Sukwan Handali, Ihsanawati, Dessy Natalia
Format: Article in Journal/Newspaper
Language:English
Published: Wolters Kluwer Medknow Publications 2017
Subjects:
DENV 2
Dengue virus
NS1 protein
Diagnostic kit
Pichia pastoris
Arctic medicine. Tropical medicine
RC955-962
Biology (General)
QH301-705.5
Arctic
Online Access:https://doi.org/10.1016/j.apjtb.2017.07.017
https://doaj.org/article/32bcbcfcd9f84998b041673f97e0d825
Description
Summary: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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