[1]邢雪,王元红,李传峰,等.羊口疮病毒F1L融合Fe蛋白的表达与鉴定[J].江苏农业学报,2020,(01):130-135.[doi:doi:10.3969/j.issn.1000-4440.2020.01.018]
　XING Xue,WANG Yuan-hong,LI Chuan-feng,et al.Expression and identification of orf virus F1L fused with Fe protein[J].,2020,(01):130-135.[doi:doi:10.3969/j.issn.1000-4440.2020.01.018]
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羊口疮病毒F1L融合Fe蛋白的表达与鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2020年01期
页码:
130-135
栏目:
畜牧兽医·水产养殖
出版日期:
2020-02-29

文章信息/Info

Title:
Expression and identification of orf virus F1L fused with Fe protein
作者:
邢雪1; 2; 王元红1; 2; 李传峰2; 缪秋红2; 曹昳1; 王桂军1; 刘光清2
(1.安徽农业大学动物科技学院，安徽合肥230036；2.中国农业科学院上海兽医研究所，上海200241)
Author(s):
XING Xue1; 2; WANG Yuan-hong1; 2; LI Chuan-feng2; MIAO Qiu-hong2; CAO Yi1; WANG Gui-jun1; LIU Guang-qing2
(1.College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;2.Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China)
关键词:
羊口疮病毒; F1L-Fe蛋白; 多克隆抗体
Keywords:
orf virus(ORFV); F1L-Fe protein; polyclonal antibody
分类号:
S855.3
DOI:
doi:10.3969/j.issn.1000-4440.2020.01.018
文献标志码:
A
摘要:
本研究克隆了羊口疮病毒安徽分离株的F1L基因，融合Fe蛋白编码基因后，插入载体pET-32a中，构建了重组质粒pET-F1L-Fe。将pET-F1L-Fe转化BL21(DE3)感受态细胞，用IPTG诱导表达重组蛋白F1L-Fe。SDS-PAGE分析结果显示，羊口疮病毒F1L-Fe基因在BL21(DE3)获得了正确表达。将大量表达的F1L-Fe蛋白进行纯化，然后免疫BALB/c鼠，制备了抗F1L蛋白的多克隆抗体。最后，以制备的多克隆抗体对F1L-Fe融合蛋白进行Western-blot检测和分析，结果表明F1L-Fe蛋白能与制备的多克隆抗体发生特异性反应，具有良好的反应原性。本研究结果为进一步研发羊口疮病毒亚单位疫苗提供了物质基础。
Abstract:
In this study, the F1L gene of orf virus(ORFV) Anhui isolated strain was cloned. After fusing with the coding gene of Fe protein, it was inserted into the expression vector pET-32a to construct recombinant plasmid pET-F1L-Fe. Subsequently, pET-F1L-Fe was transformed into BL21(DE3) competent cells, and the expression of recombinant protein F1L-Fe was induced by IPTG. The results of SDS-PAGE showed that the F1L-Fe gene was successfully expressed in BL21(DE3). Then the F1L-Fe protein was expressed in large quantities and purified. To prepare the polyclonal antibody against F1L protein, BALB/c mice were immunized with the recombinant protein. Western blot results indicated that the F1L-Fe protein reacted specifically with the prepared polyclonal antibody and showed good reactogenicity. In a word, the results of this study provide the material foundation for further developing the subunit vaccine against ORFV.

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备注/Memo

备注/Memo:
收稿日期：2019-08-07基金项目：国家重点研发计划项目（2016YFD0500108、2016YFD0501003）;上海市科技兴农创新项目[沪农科创字(2019)第3-3号]；中央级公益性科研院所基本科研业务费专项(2019JB06)作者简介：邢雪（1993-），女，安徽池州人，硕士研究生，主要从事畜禽传染病防治研究。（E-mail）2386242406@qq.com通讯作者：刘光清，（E-mail）liugq@shvri.ac.cn；王桂军，（E-mail）wgj@ahau.edu. Cn
更新日期/Last Update: 2020-03-13