Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system
Nipah virus (NiV) causes fatal respiratory illness and encephalitis in humans and animals. The matrix (M) protein of NiV plays an important role in the viral assembly and budding process. Thus, an access to the NiV M protein is vital to the design of viral antigens as diagnostic reagents. In this st...
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American Institute of Chemical Engineers
2016
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| Online Access: | http://psasir.upm.edu.my/id/eprint/54246/ http://psasir.upm.edu.my/id/eprint/54246/1/Expression%20and%20purification%20of%20the%20matrix%20protein%20of%20Nipah%20virus.pdf |
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| author | Dezfooli, Seyedehsara Masoomi Tan, Wen Siang Tey, Beng Ti Ooi, Chien Wei Hussain, Siti Aslina |
| author_facet | Dezfooli, Seyedehsara Masoomi Tan, Wen Siang Tey, Beng Ti Ooi, Chien Wei Hussain, Siti Aslina |
| author_sort | Dezfooli, Seyedehsara Masoomi |
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| collection | Online Access |
| description | Nipah virus (NiV) causes fatal respiratory illness and encephalitis in humans and animals. The matrix (M) protein of NiV plays an important role in the viral assembly and budding process. Thus, an access to the NiV M protein is vital to the design of viral antigens as diagnostic reagents. In this study, recombinant DNA technology was successfully adopted in the cloning and expression of NiV M protein. A recombinant expression cassette (baculovirus expression vector) was used to encode an N-terminally His-tagged NiV M protein in insect cells. A time-course study demonstrated that the highest yield of recombinant M protein (400–500 μg) was expressed from math formula infected cells 3 days after infection. A single-step purification method based on metal ion affinity chromatography was established to purify the NiV M protein, which successfully yielded a purity level of 95.67% and a purification factor of 3.39. The Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the purified recombinant M protein (48 kDa) was antigenic and reacted strongly with the serum of a NiV infected pig. |
| first_indexed | 2025-11-15T10:38:57Z |
| format | Article |
| id | upm-54246 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T10:38:57Z |
| publishDate | 2016 |
| publisher | American Institute of Chemical Engineers |
| recordtype | eprints |
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| spelling | upm-542462018-03-15T01:52:35Z http://psasir.upm.edu.my/id/eprint/54246/ Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system Dezfooli, Seyedehsara Masoomi Tan, Wen Siang Tey, Beng Ti Ooi, Chien Wei Hussain, Siti Aslina Nipah virus (NiV) causes fatal respiratory illness and encephalitis in humans and animals. The matrix (M) protein of NiV plays an important role in the viral assembly and budding process. Thus, an access to the NiV M protein is vital to the design of viral antigens as diagnostic reagents. In this study, recombinant DNA technology was successfully adopted in the cloning and expression of NiV M protein. A recombinant expression cassette (baculovirus expression vector) was used to encode an N-terminally His-tagged NiV M protein in insect cells. A time-course study demonstrated that the highest yield of recombinant M protein (400–500 μg) was expressed from math formula infected cells 3 days after infection. A single-step purification method based on metal ion affinity chromatography was established to purify the NiV M protein, which successfully yielded a purity level of 95.67% and a purification factor of 3.39. The Western blotting and enzyme-linked immunosorbent assay (ELISA) showed that the purified recombinant M protein (48 kDa) was antigenic and reacted strongly with the serum of a NiV infected pig. American Institute of Chemical Engineers 2016 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54246/1/Expression%20and%20purification%20of%20the%20matrix%20protein%20of%20Nipah%20virus.pdf Dezfooli, Seyedehsara Masoomi and Tan, Wen Siang and Tey, Beng Ti and Ooi, Chien Wei and Hussain, Siti Aslina (2016) Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system. Biotechnology Progress, 32 (1). pp. 171-177. ISSN 8756-7938; ESSN: 1520-6033 http://onlinelibrary.wiley.com/doi/10.1002/btpr.2192/abstract Nipah virus; Matrix protein; Baculovirus; Insect cells; Expression; Purification 10.1002/btpr.2192 |
| spellingShingle | Nipah virus; Matrix protein; Baculovirus; Insect cells; Expression; Purification Dezfooli, Seyedehsara Masoomi Tan, Wen Siang Tey, Beng Ti Ooi, Chien Wei Hussain, Siti Aslina Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title | Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title_full | Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title_fullStr | Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title_full_unstemmed | Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title_short | Expression and purification of the matrix protein of Nipah virus in baculovirus insect cell system |
| title_sort | expression and purification of the matrix protein of nipah virus in baculovirus insect cell system |
| topic | Nipah virus; Matrix protein; Baculovirus; Insect cells; Expression; Purification |
| url | http://psasir.upm.edu.my/id/eprint/54246/ http://psasir.upm.edu.my/id/eprint/54246/ http://psasir.upm.edu.my/id/eprint/54246/ http://psasir.upm.edu.my/id/eprint/54246/1/Expression%20and%20purification%20of%20the%20matrix%20protein%20of%20Nipah%20virus.pdf |