羊口疮病毒B2L基因密码子优化及截短表达
Codon Optimization and Truncated Expression of the B2L Gene of ORFV
  
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中文摘要:
      B2L蛋白是羊口疮病毒(ORFV)的结构蛋白,也是病毒的保护性抗原,可以刺激机体产生强烈的抗病毒反应。为表达出高可溶性的B2L蛋白并检测其免疫原性,将羊口疮病毒B2L基因进行密码子偏爱性优化设计,并截取抗原性高、疏水性好的区域,构建了His-tag融合可溶性标签的表达重组载体,转化至大肠杆菌BL21(DE3)感受态中,经IPTG诱导,利用SDS-PAGE电泳对融合蛋白的可溶性表达进行检测,筛选出高可溶性表达的His融合蛋白。重组蛋白通过Ni-NTA agarose亲和纯化后,进行Western-blot验证。结果显示,经过SDS-PAGE和Western- blot鉴定的目的条带分子质量与预期大小相符。本试验成功在上清中表达和纯化出B2L基因编码蛋白。
英文摘要:
      The B2L protein is a structural protein of the sheep's aphthous virus(ORFV)and is also the protective antigen,which can stimulate the body to produce strong antiviral reaction. In order to express highly soluble B2L protein and to detect the immunogenicity,the codon bias of B2L gene was optimized,then the region with high antigenicity and good hydrophobicity was intercepted,and the His-tag recombinant vector was constructed,the recombinant plasmids were transformed into E. coli BL21(DE3). After induced by IPTG,the solubility of expressed recombinant protein was tested by SDS-PAGE electrophoresis,and the highly soluble His-tag protein was screened. At last,the recombinant protein was purified by Ni-NTA agarose affinity and verified by Western-blot. The results showed the molecular weight of the band determined by SDS-PAGE and Western-blot was consistent with the expected size,hence B2L protein was successfully expressed and purified in the supernatant.
作者单位
张静,范峻豪,刘春羽,赵一,温永俊,张七斤  
中文关键词:  羊口疮病毒  B2L基因  截短表达  密码子优化
英文关键词:ORFV  B2L gene  truncated expression  codon optimization
基金项目:国家重点研发计划项目(2017YFD0500903)
DOI:10.3969/j.issn.1005-944X.2018.10.025
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