[1]吕炫,张淼,胡增金,等.鹅星状病毒衣壳纤突蛋白VP27多克隆抗体的制备和鉴定[J].江苏农业学报,2021,(02):412-417.[doi:doi:10.3969/j.issn.1000-4440.2021.02.018]
　LYU Xuan,ZHANG Miao,HU Zeng-jin,et al.Preparation and identification of polyclonal antibody against the capsid spike protein VP27 of goose astrovirus[J].,2021,(02):412-417.[doi:doi:10.3969/j.issn.1000-4440.2021.02.018]
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鹅星状病毒衣壳纤突蛋白VP27多克隆抗体的制备和鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2021年02期
页码:
412-417
栏目:
畜牧兽医·水产养殖
出版日期:
2021-04-30

文章信息/Info

Title:
Preparation and identification of polyclonal antibody against the capsid spike protein VP27 of goose astrovirus
作者:
吕炫1; 2; 张淼1; 胡增金1; 李佳明1; 张冲1; 朱英奇1; 魏娟文1; 吴琼1; 张瑞辰1; 夏伦志3; 王桂军1; 2
（1.安徽农业大学动物科技学院，安徽合肥230036；2.兽医病理生物学与疫病防控安徽省重点实验室，安徽合肥230036；3.安徽省农业科学院畜牧兽医研究所，安徽合肥230036）
Author(s):
LYU Xuan1; 2; ZHANG Miao1; HU Zeng-jin1; LI Jia-ming1; ZHANG Chong1; ZHU Ying-qi1; WEI Juan-wen1; WU Qiong1; ZHANG Rui-chen1; XIA Lun-zhi3; WANG Gui-jun
（1.College of Animal Science and Technology，Anhui Agricultural University，Hefei 230036，China;2.Anhui Provincial Key Laboratory of Veterinary Pathobiology and Disease Control，Hefei 230036，China;3.Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Sciences，Hefei 230036，China）
关键词:
鹅星状病毒; 衣壳纤突蛋白VP27; 多克隆抗体
Keywords:
goose astrovirus; capsid spike protein VP27; polyclonal antibody
分类号:
S858.332.65+9.6
DOI:
doi:10.3969/j.issn.1000-4440.2021.02.018
文献标志码:
A
摘要:
采用RT-PCR扩增获得鹅星状病毒（Goose astrovirus）的衣壳纤突蛋白基因vp27，并将其克隆至pCold-SUMO原核表达载体。重组表达载体pCold-vp27经过测序和双酶切验证正确后，转化到感受态细胞Rosetta。挑取阳性克隆子，利用异丙基硫代半乳糖苷诱导表达鹅星状病毒衣壳纤突重组蛋白，使用镍柱纯化衣壳纤突蛋白VP27，将定量的重组蛋白免疫BALB/c小鼠。利用ELISA、Western blot和IFA鉴定鼠抗衣壳纤突蛋白VP27多克隆抗体。结果显示，鼠抗衣壳纤突蛋白VP27多克隆抗体的效价大于1∶64 000，Western blot试验结果证明了鼠抗衣壳纤突蛋白VP27多克隆抗体的特异性，IFA试验结果证明鼠抗衣壳纤突蛋白VP27多克隆抗体能够识别天然的鹅星状病毒衣壳纤突蛋白VP27。
Abstract:
The vp27 gene of capsid spike protein of goose astrovirus was amplified by RT-PCR and cloned into prokaryotic expressional vector pCold-SUMO. The recombinant expressional vector pCold-vp27 was transformed into competent cell Rosetta after verified to be correct by sequencing and double enzyme digestion. The positive clones were selected, and the capsid spike recombinant protein of goose astrovirus was induced to express by isopropyl-β-D-thiogalactoside (IPTG). The capsid spike protein VP27 was purified by nickel column and quantitative recombinant protein was used to immune BALB/c mice. The polyclonal antibody against capsid spike protein VP27 in mice was identified by ELISA, Western blot and IFA. The results showed that the titer of polyclonal antibody against capsid spike protein VP27 in mice was higher than 1∶64 000. The specificity of polyclonal antibody against capsid spike protein VP27 in mice was proved by the testing results of Western blot. Results of IFA test proved that the polyclonal antibody against capsid spike protein VP27 in mice could recognize natural goose astrovirus capsid spike protein VP27.

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备注/Memo

备注/Memo:
收稿日期：2020-09-12基金项目：科技部国家重点研发计划项目（2018YFD0500100、2016YFD0500805）；安徽省家禽产业技术体系项目（2019-2020）作者简介：吕炫（1996-），男，安徽安庆人，硕士研究生，研究方向为畜禽传染病防治。(Tel)18788842820;(E-mail)961579879@qq.com通讯作者：王桂军，(E-mail)wgj@ahau.edu.cn
更新日期/Last Update: 2021-05-10