新城疫病毒调控宿主YB-1蛋白的初步研究  

Preliminary study on the regulation of host YB-1 protein mediated by Newcastle disease virus

作　　者：张世伟[1,2] 仇旭升[2] 周昌娈 孙英杰[2] 詹媛[2] 谭磊[2] 宋翠萍[2] 孟春春[2] 廖瑛[2] 茅翔[2] 丁铲[2] 王桂军[1] 

机构地区：[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

出　　处：《中国兽医科学》2016年第6期731-736,共6页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2011AA10A209);公益性行业(农业)科研专项(201303033)

摘　　要：为了揭示YB-1蛋白在新城疫病毒(NDV)感染中的作用,对NDV感染细胞中YB-1表达水平以及亚细胞定位的变化进行了初步探索。利用RT-PCR技术,从鸡胚成纤维细胞中扩增了鸡源YB-1蛋白基因,序列分析结果显示其与其他物种YB-1有较高的相似性,与人YB-1的核苷酸和氨基酸相似性分别为88.3%和78.9%。随后利用间接免疫荧光技术检测了感染NDV强毒株Herts/33的He La和DF-1细胞中内源性YB-1蛋白的亚细胞定位变化,用q RT-PCR方法检测了YB-1 m RNA表达水平,通过Western-blot检测了YB-1蛋白水平的时相变化。结果显示,He La细胞中YB-1主要存在于细胞质,而禽源DF-1细胞YB-1集中在细胞核内;感染NDV后,两种细胞中的YB-1蛋白均在细胞质内呈现颗粒化聚集,同时在NDV感染后期He La细胞中YB-1蛋白出现少量降解。该研究初步揭示了NDV感染过程中YB-1蛋白的迁移现象,推测与NDV的复制过程相关,其具体的作用机制以及对NDV复制的影响尚待更深入的研究。In order to determine a relationship between YB-1 protein and Newcastle disease virus（NDV）infection,the expression level and subcellular localization of YB-1 protein were analyzed in the NDV-infected cells.The chicken YB-1 gene was cloned from chicken embryo fibroblast（CEF） via RT-PCR.The sequence analysis showed chicken YB-1 shares similarities with YB-1 from other species.The similarity between chicken and human YB-1 gene is 88.3% in nucleotide sequence and 78.9% in amino acid sequence.Subsequently,the subcellular localization of YB-1 was determined by indirect immunofluorescent assay in the Herts/33-infected He La and DF1 cells;q RT-PCR assay and Western-blot analysis were also utilized to detect the changes of YB-1 in m RNA and protein expression levels.Our results demonstrated that YB-1protein was mainly localized in the cytoplasm of He La cells and the nucleus of DF-1 cells.The infection of NDV induced YB-1 puncta in cytoplasm.Meanwhile,the slight degradation of YB-1 was observed in He La cells.In summary,our studies showed NDV infection resulted in YB-1 translocation,suggesting that YB-1might play important roles in virus infection.Further investigations are necessary to clarify this.

关 键 词：新城疫病毒 YB-1蛋白 聚集化 降解 

分 类 号：S852.659.5[农业科学—基础兽医学]