| 猪伪狂犬病毒gB和gC蛋白B细胞表位编码序列的融合表达及活性测定 |
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| 引用本文： | 张冲,刘芳,赵东,邓瑞广,张改平,李学伍.猪伪狂犬病毒gB和gC蛋白B细胞表位编码序列的融合表达及活性测定[J].河南农业科学,2016(1):119-123. |
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| 作者姓名： | 张冲 刘芳 赵东 邓瑞广 张改平 李学伍 |
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| 作者单位： | 1. 河南科技大学 动物科学技术学院,河南 洛阳471003;河南省农业科学院动物免疫学重点实验室,河南 郑州450002;2. 河南科技大学 动物科学技术学院,河南 洛阳,471003;3. 河南省农业科学院动物免疫学重点实验室,河南 郑州,450002;4. 河南省农业科学院动物免疫学重点实验室,河南 郑州450002;河南农业大学 牧医工程学院,河南 郑州450002 |
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| 摘 要： | 依据猪伪狂犬病毒g B和g C蛋白的B细胞表位编码序列,分别设计g B和g C蛋白B细胞表位编码序列的融合扩增引物,利用融合PCR技术,将g B和g C蛋白B细胞表位编码序列有机地融合为g B-C表位编码序列。将融合基因片段插入到原核表达载体p ET28a,并转化大肠杆菌BL21(DE3),1.0 mmol/L异丙基硫代半乳糖苷(IPTG)诱导表达重组蛋白。SDS-PAGE结果显示,融合蛋白g B-C能够高效表达,重组蛋白的分子质量约为38 ku;经Western blot鉴定分析,小鼠抗His标签单克隆抗体和PRV阳性血清均能识别表达的重组蛋白。表明成功表达了融合蛋白g B-C,且表达的重组蛋白具有较好的免疫学活性。
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| 关 键 词： | 猪伪狂犬病毒 gB蛋白 gC蛋白 B细胞表位 融合表达 |
Fusion Expression and Activity Detection of Porcine Pseudorabies Virus gB and gC Glycoprotein B-cell Epitope Coding Sequences |
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| Abstract: | According to PRV B cell epitope sequences of the gB and gC protein,the fusion primers were designed respectively. Using the fusion PCR technology, the gB-C was fused by the B cell epitope se-quences of the gB and gC protein. Then the gB-C was inserted into the pET28a. The recombinant plasmid was transformed into E. coli (BL21). The E. coli (BL21)expressed the recombinant protein which was in-duced with 1 mmol/L IPTG. The result of SDS-PAGE showed that the gB-C was expressed successfully. The molecular weight of the recombinant protein was 38 ku. The result of Western blot showed that the recom-binant gB-C protein could be recognized by His-tag mouse Mcab and PRV porcine positive serum. In this research,the recombinant gB-C protein was expressed successfully and the recombinant protein had a good immunological activity. |
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| Keywords: | PRV gB protein gC protein B-cell epitope fusion expression |
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