| PCV2 ORF2与PRRSV ORF5抗原表位区串联原核表达及免疫原性 |
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| 引用本文： | 杨睿,;付利芝,;杨柳,;周雪,;沈克飞,;王孝友,;郑华,;徐登峰.PCV2 ORF2与PRRSV ORF5抗原表位区串联原核表达及免疫原性[J].兽医大学学报,2014(11):1721-1724. |
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| 作者姓名： | 杨睿 ;付利芝 ;杨柳 ;周雪 ;沈克飞 ;王孝友 ;郑华 ;徐登峰 |
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| 作者单位： | [1]重庆市畜牧科学院农业部养猪科学重点实验室,重庆402460; [2]重庆市养猪科学市级重点实验室,重庆402460 |
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| 基金项目： | 重庆市科委基本业务费资助项目（2013cstc-jbky-00903） |
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| 摘 要： | 根据GenBank公布的PCV2与PRRSV的核苷酸序列选定2个主要的抗原表位,2个抗原表位通过1个柔性的Linker相连,最后对目标片段的核苷酸序列进行修饰,使它们适合在DE3宿主菌中表达,合成目的片段729bp;将融合的核酸序列通过酶切位点BamhⅠ和XholⅠ插入pGEX-4T-1原核表达载体;用BL21（DE3）作为宿主菌,37℃下0.5mmol/L IPTG诱导表达,表达量为0.2g/L;重组蛋白免疫按80mg/头免疫仔猪,免疫后2周可使仔猪同时产生PVC2与PRRSV抗体。
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| 关 键 词： | PCV2 PRRSV 抗原 表达 |
The tandem expressing of the antigen epitope genes of PCV2-ORF2 and PRRSV- ORF5 and primary identification of its immunogenicity |
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| Institution: | YANG Rui,FU Li-zhi ,YANG Liu ,ZHOU Xue ,SHEN Ke-fei ,WANG Xiao-you ,ZHENG Hua ,XUE Deng-feng (1. Key Laboratory of Pig Industry Sciences of Agriculture Ministry, Chongqing Academy of Animal Sciences, Chongqing 402460, China ; 2. Chongqing Key Laboratory of Pig Industry Sciences, Chongqing 402460, China) |
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| Abstract: | Two major fragment of epitope of PCV2 and PRRSV were chosen from GeneBank which were connected by a flexible Linker. The nucleotide sequence of the target fragment was modified,so it was suitable for expression in host strain DE3. The purposes fragment was 729 bp. The host strain BL21 （DE3） was expressed through induced by 0.5 mmol/L IPMG in 37℃ ,the protein expressed was 0.2 g/L. The recombinant protein was used to immunize pigs,80 mg each. PCV2 and PRRSV antibodies were found in the pig at the same time after two weeks immunization. |
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| Keywords: | PCV2 PRRSV antigen expression |
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