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牛病毒性腹泻病毒P20及P14蛋白基因的表达
及其产物的抗原性分析
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李慧昕1,2，王君伟1
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（1.东北农业大学 动物医学院，黑龙江 哈尔滨150030;
2.中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室，黑龙江 哈尔滨150001 ）
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摘要：
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将牛病毒性腹泻病毒（bovine viral diarrhea virus, BVDV）p20和p14基因分别亚克隆至原核表达载体pGEX-6p-1和pPROEX-HTb，并转化至相应的宿主菌大肠杆菌BL21（DE3）pLysS和DH5α中表达。P20蛋白在2个宿主中都获得了高效表达；P14蛋白在BL21（DE3）pLysS中表达量较低，而在DH5α中未见表达。对P14蛋白诱导表达过程的监测表明，P14蛋白对大肠杆菌是一种毒性蛋白。用Western-blotting分析未能检测到两种蛋白的反应条带，推测这两种蛋白可能存在构象表位。 |
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关键词：
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牛病毒性腹泻病毒；P20蛋白；P14蛋白；原核表达；抗原性 |
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中图分类号：S 852.659.6:Q 786 文献标识码：
A 文章编号：1673-4696(2006)10-0795-05 |
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Expression of P20 and P14 protein genes of bovine viral diarrhea
virus and antigenicity analysis of the expressed proteins |
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LI Hui-xin1，2, WANG Jun-wei1 |
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(1.College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China；
2.National Key Laboratory of Vererinary Biotechnology/Harbin Veterinary Research Institute, Chinese
Academy of Agricultural Sciences, Harbin 150001, China) |
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Abstract: The P20 and P14 protein genes of bovine viral diarrhea virus (BVDV) were subcloned into two prokaryotic expression vector, pGEX-6p-1 and pPROEX-HTb, respectively, to construct recombinant expression plasmids. The recombinant plasmids were transformed into competent BL21(DE3)pLysS and DH5α accordingly for expression. Protein P20 was highly expressed in each of the host bacteria induced by IPTG. P14 protein was expressed at low level in BL21(DE3)pLysS, but not expressed in DH5α. Detection of D600nm values in the course of expression of the P14 protein showed that the P14 protein was toxic to Escherichia coli. It was proposed that these two proteins may contain conformational epitopes because no reaction was observed using Western-blotting. |
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Key words: bovine viral diarrhea virus; P20 protein; P14 protein; prokaryotic expression; antigenicity |
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