中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 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 中国预防兽医学报 2019 (12) 1204-1209
Position: Home > Articles > Identification of host proteins interacting with Clostridium perfringen β2 toxin Chinese Journal of Preventive Veterinary Medicine 2019 (12) 1204-1209

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

作  者：
曾秀;张兰桥;周姣;王婧祺;戴益民;张锦华
单  位：
江西农业大学动物科学技术学院;江西农业大学生物科学与工程学院
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)
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。
译  名：
Identification of host proteins interacting with Clostridium perfringen β2 toxin
作  者：
ZENG Xiu;ZHANG Lan-qiao;ZHOU Jiao;WANG Jing-qi;DAI Yi-min;ZHANG Jin-hua;College of Animal Science and Technology,Jiangxi Agricultural University;College of Biology Science and Engineering,Jiangxi Agricultural University;
关键词：
beta2 toxin;;Clostridium Perfringen;;IPEC-1;;yeast two-hybrid system;;LGALS1
摘  要：
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.

相似文章

导出引用
计量
文章访问数: 12
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46