人呼吸道合胞病毒RNA聚合酶复合体蛋白N、P的表达鉴定.pdfVIP

第 21 卷 3 期 中 国 病 毒 学 21 （3 ）：213-216 2006 年 5 月 VIROLOGICA SINICA May 2006 人呼吸道合胞病毒RNA 聚合酶复合体蛋白N 、P 的 表达鉴定* ** 虞结梅，何金生 ，李栋梁，杨 兵，袁 媛 （安徽医科大学免疫学教研室，安徽合肥 230032 ） Cloning and Expression of N and P of Human Respiratory Syncytial Virus RNA Polymerasa Complex ** YU Jie-mei, He Jin-sheng , LI Dong-liang, YANG Bing, YUAN Yuan Department of Immunology, Anh.ui Medical University, Hefei 230032, China ( ) Abstract ：The n and p genes from subgroup A human respiratory syncytial virus (HRSV), were amplified by RT-PCR. The n product was cloned into pGEM-T easy vector and that of p into pcDNAII vector and the two genes were subcloned into the eukaryotic expression vector pcDNA3.1(+). The resulting recombinant plasmids pcDNA3.1(+)/N and pcDNA3.1(+)/P were checked by restriction enzymes and DNA sequencing and were tranfected into COS-7 cells with the aid of Lipofectamine 2000. Expression of N and P were detected by western blots at 72 hours post transfections. Expressed N and P in COS-7 cells was confirmed by western blot. The constructed expression vectors will be used for the further reverse genetics experimentations on HRSV. Key words ：Human respiratory syncytial virus(HRSV); Nucleocapsid protein; Phosphoprotein; Eukaryotic expression 摘要：本研究根据编码 A 亚型人呼吸道合胞病毒（Human Respiratory Syncytial Virus ，HRSV ）核壳体蛋白 （