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  • Title: [Expression of tick-borne encephalitis virus prM-E protein in insect cells and studies on its antigenicity].
    Author: Liu YL, Si BY, Hu Y, Zhang Y, Yang YH, Zhu QY.
    Journal: Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi; 2005 Dec; 19(4):335-9. PubMed ID: 16415990.
    Abstract:
    BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: The prM-E protein expressed in insect cells retains good antigenicity.
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