利用PRNP--羊制备朊蛋白多克隆抗体及其特性.pdfVIP

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利用PRNP--羊制备朊蛋白多克隆抗体及其特性.pdf

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利用PRNP--羊制备朊蛋白多克隆抗体及其特性

2012,45(7):1399-1405 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.07.019 1 2 3 201210 201210; , 266032 Production and Characterization of Anti-PrP Polyclonal Antibody by PRNP Gene Knockout Goat 1 1 3 2 1 2 WANG Xue-bin , YU Hui-qing , WU Xiao-dong , XU Xu-jun , CHEN Jian-quan , CHENG Guo-xiang 1 2 ( Shanghai Transgenic Research CenterShanghai 201210; Shanghai Genon Bioengineering Limited Company, Shanghai 201210 3China Animal Health and Epidemiology Center, Qingdao 266032, Shandong) Abstract:ObjectiveThe study was conducted in order to produce and characterize anti-PrP polyclonal antibodies by prion protein knockout (PRNP-/-) goat.MethodUsing polymerase chain reaction (PCR) method, the DNA sequence encoding the goat mature PrP (Prion Protein) was amplified from goat DNA. It was then cloned into the vector pET-22b to express recombinant PrP protein. After purification, the recombinant PrP was used as antigen to immunize PRNP-/- goat for producing anti-PrP antibodies. ELISA and Western blot were conducted to characterize the titer and specificity of polyclonal antibodies.ResultA large scale of anti-PrP polyclonal antibodies was successfully produced by PRNP-/- goat. ELISA assay revealed that the titer of the polyclonal antibodies against PrP was as high as 125 600. Western blot test showed that the antibody was not