PRRSV 2b蛋白与LGALS1相互作用的验证研究  

The Validation Research on Interaction Between PRRSV 2b Protein and LGALS1

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作  者:杨达汉 邓守全 崔一龙 刘锴[1] 王学理[1] 霍晓伟[1] 马德慧[1] 薛江东[1] Yang Dahan;Deng Shouquan;Cui Yilong;Liu Kai;Wang Xueli;Huo Xiaowei;Ma Dehui;Xue Jiangdong(College of Animal Science and Technology,Inner Mongolia University for Nationalities,Tongliao Inner Mongolia 02800)

机构地区:[1]内蒙古民族大学动物科学技术学院,内蒙古通辽028000

出  处:《中国农学通报》2018年第29期112-117,共6页Chinese Agricultural Science Bulletin

基  金:国家自然科学基金项目"猪繁殖与呼吸综合征病毒2b蛋白与宿主细胞蛋白相互作用研究"(31460662);内蒙古自治区自然科学基金项目"2b蛋白介导猪繁殖与呼吸综合征病毒感染易感细胞的分子机制研究"(2017MS0319);内蒙古自治区高等学校科学研究项目"猪繁殖与呼吸综合征病毒2b蛋白介导的宿主细胞离子通道变化研究"(NJZY13163)

摘  要:旨在研究猪繁殖与呼吸综合症病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)2b蛋白与宿主细胞蛋白之间的相互作用。本研究利用酵母回返试验、GST-pull down试验、免疫共沉淀试验3种方法对LGALS1与PRRSV 2b蛋白间是否相互作用进行验证。结果表明:在酵母回返试验中,含有LGALS1与2b基因的杂交酵母菌株在SD/-Trp-Leu-His-Ade/AbA/X-α-gal的平板上生长出蓝色菌落;在GST-pull down试验中,LGALS1与2b蛋白反应的样品经Western blot可检测到融合蛋白带;在免疫共沉淀试验中,LGALS1与2b蛋白反应的样品经HA单抗沉淀后,经Western blot可检测到相应蛋白带。该研究证明了LGALS1与PRRSV 2b蛋白间发生相互作用,为研究PRRSV感染途径和增殖过程提供了理论基础。The aim is to study the interaction between porcine reproductive and respiratory syndrome virus(PRRSV) 2 b protein and host cell proteins. In this study, we used yeast regression test, GST-pull down assayand co-immunoprecipitation assay to verify the interaction between LGALS1 and PRRSV 2 b. The resultsshowed that in the yeast regression test, hybrid yeast strain of LGALS1 and 2 b gene grew blue yeast colonies onSD/-Trp-Leu-His-Ade/AbA/X-α-gal plates. In the GST-pull down assay, the fusion protein bands weredetected by Western blot in the samples reacted with 2 b protein and LGALS1. In the co-immunoprecipitationassay, samples reacted with 2 b protein and LGALS1 were precipitated with HA monoclonal antibody and thecorresponding protein bands were detected by Western blot. This paper demonstrates the interaction betweenLGALS1 and PRRSV 2 b protein and provides a theoretical basis for studying PRRSV infection andproliferation.

关 键 词:2b蛋白 LGALS1(凝集素-1) 酵母双杂交 蛋白质体外结合实验 免疫共沉淀 

分 类 号:S852.659.6[农业科学—基础兽医学]

 

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