Construction and expression of a synthetic gene encoding nonstructural glycoprotein NS1 of dengue 2 virus in Pichia pastoris
| Autor: | Riski Dwimalida Putri, Raden Roro Rika Damayanti, Bachti Alisjahbana, Aisyah, Anita Yuwita, Dessy Natalia, Sukwan Handali, Ihsanawati, Fernita Puspasari |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Antigenicity lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 medicine.drug_class viruses 030106 microbiology Dengue virus medicine.disease_cause Monoclonal antibody Biochemistry Genetics and Molecular Biology (miscellaneous) NS1 protein law.invention Pichia pastoris 03 medical and health sciences Affinity chromatography law medicine lcsh:QH301-705.5 chemistry.chemical_classification Expression vector biology Chemistry virus diseases biochemical phenomena metabolism and nutrition biology.organism_classification Virology 030104 developmental biology lcsh:Biology (General) Biochemistry DENV 2 Diagnostic kit Recombinant DNA Glycoprotein |
| Zdroj: | Asian Pacific Journal of Tropical Biomedicine, Vol 7, Iss 8, Pp 689-693 (2017) |
| ISSN: | 2221-1691 |
| DOI: | 10.1016/j.apjtb.2017.07.017 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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