Expression and Identification of F Protein of Peste Des Petits Ruminants Virus Strain Tibet 07 in Insect Cells |
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Authors: | ZHAO Wen-nian WANG Qing-hua LIU Chun-ju WANG Shu-juan LEI Cheng-hong BAO Jing-yue WANG Zhi-liang |
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Institution: | 1. China Animal Health and Epidemiology Center, Qingdao 266032,China;2. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830000, China |
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Abstract: | In order to obtain purified peste des petits ruminants virus F protein, the F gene of peste des petits ruminants virus strain Tibet 07 was cloned into transfer vector pFastBac/CT-TOPO, plasmid pFastBacCT-PPRV-F was then transformed into DH10Bac complement cells. The recombinant bacmid DNA was isolated and transfected into sf9 insect cells to express F protein. The expressed F protein was identified by using SDS-PAGE, Western blotting and indirect immunofluorescence assay. The recombinant F protein was successfully expressed in insect cells with a relative molecular of 59 ku.The sf9 cells infected by Bacmid-PPRV-F can reacted with positive serum of peste des petits ruminants.Using the Bac-to-Bac baculovirus expression system expressed the F protein of peste des petits ruminants virus strain Tibet 07.This work provides basis for development of rapid test for detection of peste des petits ruminants antibody. |
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Keywords: | peste des petits ruminants F protein baculovirus expression system |
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