| 牛病毒性腹泻病毒E0基因的原核表达与活性检测 |
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| 引用本文： | 严伟行.牛病毒性腹泻病毒E0基因的原核表达与活性检测[J].中国畜牧兽医,2013,40(1):54-56. |
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| 作者姓名： | 严伟行 |
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| 作者单位： | 青海省西宁市湟中县上新庄镇兽医站, 青海西宁 811602 |
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| 摘 要： | 参考牛病毒性腹泻病毒(bovine viral diarrhea virus,BVDV)基因组序列设计1对引物,扩增出650 bp的E0基因片段。将目的片段克隆至pMD18-T载体,经酶切鉴定获得阳性重组质粒并对其进行测序。将E0基因定向亚克隆到pET32a表达载体中,酶切及测序鉴定正确后,转化BL21表达菌,经IPTG诱导得到了以包涵体形式表达的重组蛋白。重组蛋白经亲和层析法纯化后,免疫印迹检测证明纯化的重组蛋白具有良好的活性。
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| 关 键 词： | 牛病毒性腹泻病毒 E0基因 原核表达 活性检测 |
| 收稿时间： | 2012-06-27 |
Expression and Characterization of E0 Protein of Bovine Viral Diarrhea Virus |
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| YAN Wei-hang.Expression and Characterization of E0 Protein of Bovine Viral Diarrhea Virus[J].China Animal Husbandry & Veterinary Medicine,2013,40(1):54-56. |
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| Authors: | YAN Wei-hang |
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| Institution: | Veterinary Station, Shangxinzhuang Town, Huangzhong County of Qinghai Province, Xining 811602, China |
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| Abstract: | A fragment of about 650 bp was amplified by RT-PCR technique with specific primers based on BVDV genome sequence. Then the target fragment was directionally cloned into pET32a vector. After identifying with enzyme cut and sequencing, the recombinant plasmid was transformed into E. coli BL21(DE3). The recombinant protein E0 was expressed in inclusion body form in E. coli after induction with IPTG. After purification, the purified protein was analyzed by Western blotting, the results showed that the purified recombinant protein retained better antigenicity. |
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| Keywords: | bovine viral diarrhea virus E0 gene prokaryotic expression characterization |
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