[1]曾 秀,张兰桥,周 姣,等.猪源A型产气荚膜梭菌β2毒素与宿主细胞互作蛋白的筛选鉴定[J].中国预防兽医学报,2019,(12):1204-1209.[doi:10.3969/j.issn.1008-0589.201905010]
 ZENG Xiu,ZHANG Lan-qiao,ZHOU Jiao,et al.Identification of host proteins interacting with Clostridium perfringen β2 toxin[J].Chinese journal of preventive veterinary medicine,2019,(12):1204-1209.[doi:10.3969/j.issn.1008-0589.201905010]
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猪源A型产气荚膜梭菌β2毒素与宿主细胞互作蛋白的筛选鉴定()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年12
页码:
1204-1209
栏目:
病原生物学
出版日期:
2020-01-25

文章信息/Info

Title:
Identification of host  proteins interacting with Clostridium perfringen β2 toxin
文章编号:
1008-0589(2019)12-1204-06
作者:
 

曾 秀1张兰桥2周 姣1王婧祺1戴益民1张锦华1*

 (1. 江西农业大学 动物科学技术学院,江西 南昌 330045;2. 江西农业大学 生物科学与工程学院,江西 南昌 330045)
Author(s):
 

ZENG Xiu1 ZHANG Lan-qiao2 ZHOU Jiao1 WANG Jing-qi1 DAI Yi-min1 ZHANG Jin-hua1*

 (1. College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, China;
2. College of Biology Science and Engineering, Jiangxi Agricultural University,  Nanchang 330045, China)
关键词:
β2毒素产气荚膜梭菌IPEC-1酵母双杂交系统半乳糖凝集素(LGALS1)
Keywords:
 beta2 toxin  Clostridium Perfringen  IPEC-1  yeast two-hybrid system  LGALS1
DOI:
10.3969/j.issn.1008-0589.201905010
文献标志码:
A
摘要:
产气荚膜梭菌产生的β2毒素在仔猪肠炎中起着重要作用,为研究该毒素侵入宿主细胞的作用机制,本研究以β2为诱饵蛋白,将其与酿酒酵母转录因子 GAL4 的BD (DNA binding domain)融合表达,利用猪肠上皮细胞系IPEC-1的cDNA文库与GAL4 的AD 区(Activation domain)融合表达,通过酵母双杂交系统筛选鉴定与β2毒素相互作用的猪肠上皮细胞蛋白。筛选得到的阳性转化子经提取质粒和测序后,比对其所包含的编码蛋白序列,得到4个潜在的与β2毒素存在相互作用的细胞蛋白,分别为S20、LGALS1、L12和Atrophin-1。其中S20、L12和Atrophin-1是位于细胞内的核蛋白,而LGALS1是一种存在于细胞膜外侧的半乳糖凝集素,可能是毒素的作用位点。对LGALS1进行回交验证,结果显示其C端与β2毒素的相互作用强于其完整蛋白与β2毒素的相互作用。本研究结果对研究β2毒素的致病机制奠定了基础。
  
Abstract:
Beta2(β2) toxin of Clostridium perfringens plays an important role in piglet enteritis. In order to study the pathogenic mechanisms of this toxin, yeast two-hybrid technology was used in this study to screen and identify host proteins interacting with β2 toxin. Used as a bait protein, β2 was fused with BD domain(DNA binding domain) of the Saccharomyces cerevisiae transcription factor GAL4 for expression. The cDNA library of porcine intestinal epithelial cell line IPEC-1 was constructed, then the cDNA library was fused with AD domain of GAL4 for expression. The proteins that exist in the pig intestinal epithelial cells and interact with β2 toxin were screened and identified by yeast two-hybrid system. The positive yeast transformants were selected, and after plasmids extraction and sequencing, four potential proteins interacting with β2 toxin were obtained by comparing the sequences of the encoded proteins, namely S20, LGALS1, L12 and Atrophin-1. Among these, S20, L12 and Atrophin-1 are intracellular nucleoproteins, while LGALS1 is one kind of galectins that are located in the cell outer membrane and may be related to the invasion of toxins. Therefore, LGALS1 was verified by yeast retest analysis. The results showed that the interaction between C-terminal and β2 toxin is stronger than that of the full-length protein. The results of this experiment will play an important role in elucidating the pathogenesis of β2 toxin.

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     (本文编辑:李 娜;英文编辑:蔡文通)

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更新日期/Last Update: 2020-01-19