Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of

Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of

dengue 2 virus in Pichia pastoris

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

【期刊名称】《亚太热带生物医学杂志（英文版）》 【年(卷),期】2017(007)008

【摘要】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 af-finity chromatography, and its antigenicity was tested. Results: The recombinant DENV-2 NS1 protein was secreted as a protein with a mo-lecular weight of ～45 kDa, and the optimal expression condition was achieved by in-duction 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. 【总页数】5页(689-693)