利用酵母双杂交系统在人脑cDNA文库中筛选与禽流感.PDFVIP

生 物 工 程 学 报 Chin J Biotech 2010, August 25; 26(8): 1037−1041 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ ©2010 CJB, All rights reserved. 动物及兽医生物技术 利用酵母双杂交系统在人脑cDNA 文库中筛选与禽流感 病毒核蛋白相互作用的蛋白质 尹文偲，胡勇，金梅林 华中农业大学动物医学院 农业微生物学国家重点实验室，武汉 430070 摘 要: 禽流感病毒核蛋白 (NP) 在病毒的转录、复制以及决定病毒的宿主特异性方面都具有重要作用。通过酵母双 杂交系统筛选与核蛋白相互作用的蛋白，为进一步了解NP 蛋白与细胞内蛋白质的相互关系以及流感病毒与宿主的相互 关系奠定基础。应用酵母双杂交系统，构建NP 诱饵质粒，进而筛选人脑cDNA 文库，寻找可能与禽流感病毒NP 相互 作用的蛋白质。经过酵母双杂交共验证，得到7 个与NP 相互作用的阳性克隆。该结果为深入了解病毒复制的分子机理 及其在蛋白质水平上与宿主蛋白的相互作用关系提供了线索。 关键词: 禽流感病毒核蛋白，人脑 cDNA 文库，酵母双杂交系统，蛋白间相互作用 Screening of proteins interacting with avian influenza virus Nucleoprotein by yeast two-hybrid system in human brain cDNA library Wensi Yin, Yong Hu, and Meilin Jin State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China Abstract: Avian influenza virus Nucleoprotein (NP) is important in viral transcription, replication and determining host specificity of influenza virus. Yeast two-hybrid technique was applied to screen for proteins interacting with virus nucleoprotein, so as to further elucidate the interaction between virus nucleoprotein and cellular proteins, as well as the interaction between virus and host. To explore new proteins interacted with NP protein, a human brain cDNA library was screened using yeast two-hybrid system with NP as the bait. DNA inserts of the positive AD/library plasmids were sequenced. By the BLAST analysis against the GenBank databases seven positive clones resulted in seven genes. Our results could help for the further study on the molecular mechanism of virus replication, transcription and protein-protein interactio