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《Chinese Journal of Veterinary Science》 2001-02
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Cloning and Expression of Glycoprotein I Gene of Marek′s Disease Virus Strain G2

DING Jia-bo 1,CUI Zhi-zhong 2* ,WEI Ping 3,LU Yin-hua 4,ZHAO Wen-ming 5,HAN Ling-xia 1, JI Rong 1 (1.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China; 2.College of Animal Science and Technology,Shandong Agricultural  
Glycoprotein I(gI) gene was amplified from genomic DNA of MDV G2 with the dismission of 165 bp at the N-terminal of ORF by polymerase chain reaction (PCR) technigue. PCR product was cloned into the expressing plasmid vector pGEX-6p-1 via restriction endonucleases BamHⅠ and SmaⅠ. The recombinant was verified by restriction endonuclease analysis and nucleotide sequencing. Then it was transformed into its host E.coli strain BL 21 for gI expression. The expression of gI gene was indentified by SDS-PAGE and Western-blotting, and found to be 63 000 in size as a fussion protein with glutathione transferase protein. The specific bands of expression was excised from the gel and injected into the mice once a week and for 5 weeks. The antisera was collected from the immunized mice and used for fluorescence assay(FA) with CEF monolayers infected by RB1B and GA of MDV. The positive stainings were found in the MDV plaques and localized on the cytomembrane of the infected cells. The results showed that the in vitro expressed protein of gI gene via recombinant plasmid vector maintains some antigenicity of MDV.
【Fund】: 国家“8 6 3”计划资助项目 !(10 1-5 2 -0 3-0 2 )
【CateGory Index】: S852.65;Q78
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