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首页> 外文会议>2011 International Conference on Remote Sensing, Environment and Transportation Engineering >Prokaryotic expression of the antigenic site B/C of S gene of porcine transmissible gastroenteritis virus and preparation of its monoclonal antibodies
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Prokaryotic expression of the antigenic site B/C of S gene of porcine transmissible gastroenteritis virus and preparation of its monoclonal antibodies

机译：猪传染性胃肠炎病毒S基因抗原位点B / C的原核表达及其单克隆抗体的制备

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In this study, the antigenic site B/C of S gene of transmissible gastroenteritis virus (TGEV) was cloned into prokaryotic expression vector pET-28a and the recombinant plasmid was transferred into E.coli BL21 (DE3). The fusion protein was expressed after induction with IPTG. The SDS-PAGE purified protein was used as immunogen. Six-week-old BALB/c mice were immunized three times with recombinant protein. The hybridoma cell strain, designated 5G, steadily secreting monoclonal antibodies (McAbs) against the protein of site B/C, was obtained by hybridoma technique. It belonged to IgM subclass and could recognize the site B/C recombinant protein of S gene of TGEV. It was confirmed that the McAbs only could react with TGEV using indirect immunofluorescence assay (IFA) and indirect ELISA, indicating the McAbs had good specificity.
机译：在这项研究中，将传染性胃肠炎病毒（TGEV）S基因的抗原位点B / C克隆到原核表达载体pET-28a中，并将重组质粒转移到大肠杆菌BL21（DE3）中。用IPTG诱导后表达融合蛋白。 SDS-PAGE纯化的蛋白质用作免疫原。用重组蛋白免疫六周大的BALB / c小鼠三遍。通过杂交瘤技术获得了稳定地分泌针对B / C位点蛋白的单克隆抗体（McAbs）的杂交瘤细胞株，命名为5G。它属于IgM亚类，可以识别TGEV S基因的位点B / C重组蛋白。使用间接免疫荧光法（IFA）和间接ELISA证实了单克隆抗体只能与TGEV反应，表明该单克隆抗体具有良好的特异性。

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