| 猪圆环病毒2型Cap蛋白的截短表达与活性检测 |
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| 引用本文： | 曹贵忠.猪圆环病毒2型Cap蛋白的截短表达与活性检测[J].中国畜牧兽医,2012,39(8):91-94. |
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| 作者姓名： | 曹贵忠 |
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| 作者单位： | 青海省西宁市湟源县畜牧兽医站,青海西宁 812000 |
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| 摘 要： | 本研究参照已发表的PCV2基因组序列,设计合成1对特异性引物,以PCV2基因组DNA为模板,PCR扩增了长约480 bp的ORF2基因片段。将目的片段定向克隆到pGEX-6p-1原核表达载体,酶切及测序鉴定正确后,转化BL21(DE3)表达菌,经IPTG诱导得到了以包涵体形式表达的重组蛋白。采用亲和层析法在变性的条件下纯化重组蛋白,纯化的重组蛋白浓度为0.396 mg/mL。纯化蛋白经免疫印迹、间接ELISA检测证明具有良好的抗原性和特异性。
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| 关 键 词： | 猪圆环病毒2型 Cap蛋白 原核表达 活性检测 |
| 收稿时间： | 2012-03-22 |
Characterization of Porcine Circovirus Type 2 Truncated Cap Protein Expressed in E.coli |
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| CAO Gui-zhong.Characterization of Porcine Circovirus Type 2 Truncated Cap Protein Expressed in E.coli[J].China Animal Husbandry & Veterinary Medicine,2012,39(8):91-94. |
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| Authors: | CAO Gui-zhong |
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| Institution: | The Station of Animal Husbandry and Veterinary of Huangyuan County, Xining City in Qinghai Province, Xining 812000,China |
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| Abstract: | Designing and synthesizying one pair of primers according to the published PCV2 genome sequence,a fragment of about 480 bp long was amplified by RT-PCR technique with specific primers.Then the target fragment was directionally cloned into pGEX-6p-1 vector.After identifying with enzyme cut and sequencing,the recombinant plasmid was transformed into E.coli BL21(DE3).The recombinant Cap protein was expressed in inclusion body form after induction with IPTG.After denaturation,renaturation of recombinant protein by affinity chromatography purification,purification of the recombinant protein concentration was 0.396 mg/mL.Western blotting and indirect ELISA testing showed that the purified recombinant protein retained better antigenicity and specificity. |
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| Keywords: | porcine circovirus type 2 Cap protein prokaryotic expression activity detection |
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