Expression of tick-borne encephalitis virus prM-E protein in insect cells and studies on its antigenicity / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 335-339, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-333011
ABSTRACT
<p><b>BACKGROUND</b>To express the prM-E protein in Sf9 cells, and lay a basis for further study on the function of the viral proteins and development of specific diagnostic reagents.</p><p><b>METHODS</b>The recombinant prM-E protein of tick-borne encephalitis virus was expressed in insect cell Sf9 by RT-PCR amplification of prM-E gene, construction of donor plasmid of Bac-to-Bac baculovirus expression system, homologous recombination of donor plasmid with bacmid DNA at the site of Tn7 and transfection of insect cell Sf9.</p><p><b>RESULTS</b>Recombinant subviral particles, about 30 nm in diameter, consisting of prM-E were observed by electron microscope in the supernatant of infected cells, which indicated that infected cells released virus-like particles (VLPs) into the culture medium. The results of Western-blot and the indirect immunofluorescence assay (IFA) showed that the recombinant proteins retained antigenic and conformational structures similar to those of native virus proteins. Using the recombinant prM-E as antigens to detect samples of patient sera by ELISA and IFA, all of 16 sera from patients with tick-borne encephalitis were positive and all of 6 sera from other patients were negative.</p><p><b>CONCLUSION</b>The prM-E protein expressed in insect cells retains good antigenicity.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Gene Expression Regulation, Viral
/
Cell Line
/
Viral Envelope Proteins
/
Blotting, Western
/
Spodoptera
/
Fluorescent Antibody Technique, Indirect
/
Reverse Transcriptase Polymerase Chain Reaction
/
Encephalitis Viruses, Tick-Borne
/
Allergy and Immunology
/
Genetics
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2005
Type:
Article
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