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猪细胞因子信号传导抑制因子1蛋白的表达纯化与多克隆抗体制备 被引量:2
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作者 夏爽飞 陈鑫鑫 +4 位作者 乔松林 柴书军 罗雪刚 谢莎 张改平 《中国兽医科学》 CAS CSCD 北大核心 2019年第6期707-714,共8页
为制备猪细胞因子信号传导抑制因子1(SOCS1)的多克隆抗体,采用RT-PCR扩增猪SOCS1基因序列,将其克隆至原核表达载体pET-28a(+),构建重组表达载体pET28a-pSOCS1,并转化至大肠杆菌BL21(DE3)感受态细胞中,在不同温度、IPTG浓度、诱导时间等... 为制备猪细胞因子信号传导抑制因子1(SOCS1)的多克隆抗体,采用RT-PCR扩增猪SOCS1基因序列,将其克隆至原核表达载体pET-28a(+),构建重组表达载体pET28a-pSOCS1,并转化至大肠杆菌BL21(DE3)感受态细胞中,在不同温度、IPTG浓度、诱导时间等条件下进行表达,确定最佳表达条件,并对重组蛋白pSOCS1进行纯化。以表达的重组蛋白pSOCS1免疫新西兰大白兔制备多克隆抗体,经4次免疫,间接ELISA测得抗体效价达到1∶51 200。Western-blot和间接免疫荧光试验结果表明,该多克隆抗体可与pSOCS1重组蛋白和经HEK293T细胞过表达的pSOCS1蛋白发生特异性反应,证明pSOCS1重组蛋白免疫原性良好。本研究为后续制备单克隆抗体、研究猪SOCS1相关作用机制奠定了基础。 展开更多
关键词 猪SOCS1 生物信息学分析 原核表达 多克隆抗体
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Development of a Synthetic Peptide ELISA Assay for the Detection of Foot-and-Mouth Disease Virus Nonstructural Protein Antibodies 被引量:1
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作者 YANG Ji-fei YANG Su-zhen +7 位作者 YANG Yan-yan ZHI Ai-min ZHAO Dong ZHI Yu-bao XING Guang-xu DENG Rui-guang chai shu-jun ZHANG Gai-ping 《Agricultural Sciences in China》 CAS CSCD 2010年第11期1677-1683,共7页
In order to develop an ELISA assay with synthetic peptides for the detection of antibody to the nonstructural proteins of foot-and-mouth disease virus, specific peptides were synthesized by a solid-phase method accord... In order to develop an ELISA assay with synthetic peptides for the detection of antibody to the nonstructural proteins of foot-and-mouth disease virus, specific peptides were synthesized by a solid-phase method according to FMDV NSPs B-cell epitopes, and were conjugated to carrier protein BSA. An ELISA system was developed to detect FMDV NSPs antibody with the conjugated proteins as the coating antigen. The optimal coating concentration of the antigen was determined as 2.5 μg mL-1. The comparative study of this assay with UBI NSP ELISA kit and national commercial 3ABC ELISA kit in the detection of 199 serum samples showed that they were very coincident, and the identity rates were 96.48 and 97.48%, respectively. The development of ELISA using the synthetic peptides as coating antigen is specific, reproducible, stable, and easy, and can be used to differentiate FMDV infected pigs from immunized pigs. 展开更多
关键词 foot and mouth disease virus (FMDV) ELISA synthetic peptide
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