LIU Jian-ling, ZHANG Yan-ming, SU Zheng-yuan and XU Xin-gang. Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein[J]. Virologica Sinica, 2006, 21(3): 249-252.
Citation:
LIU Jian-ling, ZHANG Yan-ming, SU Zheng-yuan, XU Xin-gang.
Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein .VIROLOGICA SINICA, 2006, 21(3)
: 249-252.
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摘要
利用DNA重组技术将猪瘟病毒(Classical swine fever virus , CSFV)石门株囊膜蛋白E2基因插入逆转录病毒载体pBABE-puro 中构建成重组逆转录病毒载体pBABE-puro-E2,该重组逆转录病毒载体与pVSVg质粒经磷酸钙共转染法将其转入293GP细胞中包装逆转录病毒假病毒。用包装的假病毒感染PK-15细胞,经嘌呤霉素筛选阳性细胞后进行流式细胞技术(FACS)分析,结果表明CSFV E2基因在PK-15细胞膜上成功表达。将表达E2蛋白的PK-15细胞腹腔免疫Balb/c小鼠,成功诱导小鼠产生了抗E2蛋白的抗体。
Expression of CSFV E2 Envelope Protein Gene in Eukaryotic Cells and Preparation of Monoclonal Antibodies against CSFV E2 Envelope Protein
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1.
1.College of Animal Science and Technology , Northwest A & F University , Yangling , Shaanxi 712100 , China
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2.
College of Life Sciences, Northwest University, Xian 710069,China
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Corresponding author:
ZHANG Yan-ming,
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Abstract
The recombinant retroviral vector pBABE-puro-E2 was constructed by inserting full-length cDNA of CSFV Shimen strain E2 gene into pBABE-puro. Both the recombinant retroviral vector and pVSVg plasmid were transfected into eukaryotic cells 293GP by calcium phosphate transfection method, and the pseudovirus were produced. The pseudovirus-infected eukaryotic cells PK-15 and expression of E2 protein were determined by puromycin-resistant and FACS analysis. Balb/c mice were intraperitoneal injected with PK-15 cells expressing the CSFV E2 protein. Anti-CSFV E2 antibody was screened by ELISA. The results showed that CSFV E2 protein was expressed in PK-15 cells’ envelope protein successfully. ELISA could detect specific anti-E2 antibody in mouse serum immuned by PK-15 cells.
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References
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Proportional views
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