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牦牛和双峰驼重组朊蛋白的原核细胞表达
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吴润1,2，陈豪泰1，刘湘涛1，谢庆阁1
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(1. 中国农业科学院 兰州兽医研究所 农业部畜禽病毒学重点实验室，甘肃 兰州730046；
2. 甘肃农业大学 动物医学院，甘肃 兰州730070)
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摘要：
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应用重组DNA技术构建了牦牛和双峰驼重组朊蛋白的4个表达载体，即pGEXBoPrP(23～242)、pGEXBoPrP(100～242)、pGEXCaPrP(25～241)和pGEXCaPrP(93～241)，经转化E.coli BL21(DE3)，成功地获得了重组菌E.coli BoPrP(23～242)、E.coli BoPrP(100～242)、E.coli CaPrP(25～241)和E.coliCaPrP(93～241)，分别可表达大小约为50.2、41.7、49.9和42.4ku的融合蛋白。各重组菌经1.0mmol/L的IPTG于37℃诱导5～6h可产生占细菌总蛋白量30%～45%的融合蛋白。免疫印迹分析表明，除融合蛋白GSTBoPrP(100～242)外，GSTBoPrP(23～242)、GSTCaPrP(25～241)和GSTCaPrP(93～241)均可与抗牛单克隆抗体4C11反应，显示中国黄牛PrPC与牦牛和双峰驼PrPC具有共同的抗原成分。 |
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关键词：
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朊粒； 牦牛； 双峰驼； 原核细胞表达 |
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中图分类号：S 852.659.7: Q 786 文献标识码：
A 文章编号：1673-4696(2006)01-0001-06 |
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Prokaryotic expression of recombinant prion proteins of
domestic yak and Camelus bactrianus |
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WU Run1,2，CHEN
Haotai1，LIU Xiangtao1，XIE
Qingge1 |
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(1. Key Laboratory of Animal Virology, Ministry of Agriculture/Lanzhou Veterinary Research Institute，Chinese
Academy of Agricultural Sciences, Lanzhou 730046,China; 2.College of Veterinary Medicine, Gansu
Agricultural University, Lanzhou 730070,China)
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Abstract: By using the recombinant DNA technology, four expression plasmids, pGEXBoPrP(23-242), pGEXBoPrP(100-242), pGEXCaPrP(25-241) and pGEXCaPrP(93-241), of yak and camel recombinant prion proteins were constructed, After transforming E.coli BL21(DE3), the recombinant bacteria E.coli BoPrP(23-242),E.coli BoPrP (100-242),E.coli CaPrP(25-241) and E.coli CaPrP(93-241) were obtained. While the recombinant bacteria were induced by 1.0mmol/L by IPTG at 37℃ for 5h to 6 h, the expectant fusion proteins with molecular sizes of approximately 50.2ku, 41.7ku, 49.9ku and 42.4ku were produced, respectively, and the expression products accounted for 30%-45% of overall bacterium protein. Westernblotting analysis confirmed that the fusion proteins, GSTBoPrP(23-242), GSTCaPrP(25-241) and GSTCaPrP(93-241) could react with the antibovine PrP monoclonal antibody 4C11, with the exception of GSTBoPrP(100-242). Results showed that the yak PrP and the camel PrP shared the same antigen component as Chinese yellow cattle PrPC. |
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Key words: prion； domestic yak； Camelus bactrianus； prokaryotic expression |
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