This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Eshaghi, M.
Right arrow Articles by Yusoff, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Eshaghi, M.
Right arrow Articles by Yusoff, K.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, July 2005, p. 3172-3177, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3172-3177.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Purification and Characterization of Nipah Virus Nucleocapsid Protein Produced in Insect Cells

Majid Eshaghi,1 Wen Siang Tan,1,2 Swee Tin Ong,1 and Khatijah Yusoff1,2*

Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences,1 Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia2

Received 1 October 2004/ Returned for modification 5 November 2004/ Accepted 7 February 2005

The nucleocapsid (N) protein of Nipah virus (NiV) is a major constituent of the viral proteins which play a role in encapsidation, regulating the transcription and replication of the viral genome. To investigate the use of a fusion system to aid the purification of the recombinant N protein for structural studies and potential use as a diagnostic reagent, the NiV N gene was cloned into the pFastBacHT vector and the His-tagged fusion protein was expressed in Sf9 insect cells by recombinant baculovirus. Western blot analysis of the recombinant fusion protein with anti-NiV antibodies produced a band of approximately 62 kDa. A time course study showed that the highest level of expression was achieved after 3 days of incubation. Electron microscopic analysis of the NiV recombinant N fusion protein purified on a nickel-nitrilotriacetic acid resin column revealed different types of structures, including spherical, ring-like, and herringbone-like particles. The light-scattering measurements of the recombinant N protein also confirmed the polydispersity of the sample with hyrdrodynamic radii of small and large types. The optical density spectra of the purified recombinant fusion protein revealed a high A260/A280 ratio, indicating the presence of nucleic acids. Western blotting and enzyme-linked immunosorbent assay results showed that the recombinant N protein exhibited the antigenic sites and conformation necessary for specific antigen-antibody recognition.

* Corresponding author. Mailing address: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM 43400, Serdang, Selangor, Malaysia. Phone: 603 89466701. Fax: 603 89430913. E-mail: kyusoff{at}putra.upm.edu.my.

Journal of Clinical Microbiology, July 2005, p. 3172-3177, Vol. 43, No. 7
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.7.3172-3177.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Yu, F., Khairullah, N. S., Inoue, S., Balasubramaniam, V., Berendam, S. J., Teh, L. K., Ibrahim, N. S. W., Abdul Rahman, S., Hassan, S. S., Hasebe, F., Sinniah, M., Morita, K. (2006). Serodiagnosis Using Recombinant Nipah Virus Nucleocapsid Protein Expressed in Escherichia coli.. J. Clin. Microbiol. 44: 3134-3138 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»