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

Eshaghi, Majid and Tan, Wen Siang and Ong, Swee Tin and Yusoff, Khatijah (2005) Purification and Characterization of Nipah Virus Nucleocapsid Protein Produced in Insect Cells. Journal of Clinical Microbiology, 43 (7). pp. 3172-3177. ISSN 0095-1137

Full text not available from this repository.

Official URL: http://dx.doi.org/10.1128/JCM.43.7.3172-3177.2005

Abstract

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 a diagnostic reagent, the NiV n gene was cloned into the pFastBacHT vector and and his tagged fusion protian 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. At time course study showed that the highest level of expression was achived after 3 days of incubation. Electron microscopic analysis of the NiV recombinant N fusion protein purified on a nickel- nitrilotriacetic asid resin column revealed different types of structures, including spherical, ring-like, particles. The light-scattering measurements of the recombinant N protein also confirmed the polydispersity of the sample with hydrodynamic radii of small and large types. The optical density spectra of the purified recombinant fusion protein reavealed a high A 260 / A 280 ratio, indicating the presence of nucleic acids. Western blotting and enzyme-linked immunosorbent assay result showed that the recombinant N protein exhibited the antigenic sites and conformation necessary for specific antigen-antibody recognition.

Item Type:Article
Faculty or Institute:Faculty of Biotechnology and Biomolecular Sciences
Publisher:American Society for Microbiology
DOI Number:10.1128/JCM.43.7.3172-3177.2005
Altmetrics:http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1128/JCM.43.7.3172-3177.2005
ID Code:699
Deposited By:INVALID USER
Deposited On:24 Nov 2008 17:55
Last Modified:30 Jul 2010 04:21

Repository Staff Only: Edit item detail

Document Download Statistics

This item has been downloaded for since 24 Nov 2008 17:55.

View statistics for "Purification and Characterization of Nipah Virus Nucleocapsid Protein Produced in Insect Cells "


Universiti Putra Malaysia Institutional Repository

Universiti Putra Malaysia Institutional Repository is an on-line digital archive that serves as a central collection and storage of scientific information and research at the Universiti Putra Malaysia.

Currently, the collections deposited in the IR consists of Master and PhD theses, Master and PhD Project Report, Journal Articles, Journal Bulletins, Conference Papers, UPM News, Newspaper Cuttings, Patents and Inaugural Lectures.

As the policy of the university does not permit users to view thesis in full text, access is only given to the first 24 pages only.