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《Chinese Journal of Preventive Veterinary Medicine》 2001-01
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Cloning and Expression of Glycoprotein I Gene of Marek′s Disease Virus Strain 648

DING Jiabo 1,CUI Zhizhong 2,XU Jiansheng 1,WEI Ping 3,LU Yinhua 1, ZHAO Wenming 1,HAN Lingxia 1,JI Rong 1 (1.Vet.Med.coll.,Yangzhou University,Yangzhou 225009; 2.Anim.Sci.and tech.Coll.,Shangdong Agri.University,Taian 271018; 3.Dept.of Vet.Me  
Glycoprotein I(gI) gene was amplified from genomic DNA of MDV 648 with the dismission of 165bps at the N-terminal of ORF by polymerase chain reaction (PCR) technique. 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 Westernblotting, and found to be 63kD in size as a fusion protein with glutathione stransferase(GST) 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 stains were found in the MDV plaques and localized on the cytomembrane of the infected cells. The results of the study showed that the vitro expressed protein of gI gene via recombinant plasmid vector maintains some antigenicity of MDV.
【Fund】: 国家“86 3计划资助项目! (课题号 :10 1_5 2_0 3_0 2 )
【CateGory Index】: S855;;S858.31
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