猪细小病毒SY-99株非结构蛋白NS1基因的克隆及序列分析  

Cloning and Sequencing of Non-structural Protein NS1 Gene of Porcine Parvovirus SY-99 Strain

作　　者：吴丹[1] 仇华吉[1] 李昌文[1] 薛强[1] 周艳君[1] 李景鹏[2] 韩孝成[1] 刘慧敏 童光志[1] 

机构地区：[1]中国农科院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]东北农业大学生物工程系,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2002年第4期241-244,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：猪细小病毒 (Porcineparvovirus,PPV)SY_99株由本室分离 ,提取SY_99株的基因组DNA ,利用PCR扩增出全长NS1基因 ,将该基因克隆到原核表达载体pET2 8a中并测序。结果表明 ,NS1基因全长 1989bp ,编码 6 6 2个氨基酸。氨基酸序列中含有在PPV复制过程中发挥重要作用的保守基序GKRN ,并有 3个潜在的糖基化位点 ,分别位于 35 6～ 35 9、4 4 6～ 4 4 9、5 13～ 5 16位氨基酸处。SY_99株NS1基因与其它PPV毒株NADL2_1、NADL2_2、Kresse株核苷酸同源性分别为 98%、99%、99% ,氨基酸同源性分别为 97%、99%、99%。Porcine parvovirus(PPV) SY_99strain was isolated by our laboratory.We extracted the genomic DNA of SY_99 strain and amplified NS1 gene.NS1 gene was cloned into pET28a,a prokaryotic expression vector,and then sequenced.NS1 gene is 1989 bp in length and encodes 662aa.NS1 protein of SY_99 contains the conserved sequence GKRN that is importeant in replication of PPV.There are three potential N_glycosylation sites in NS1 protein of SY_99 and locate in 356_359aa,446_449aa and 513_516aa, respectively.NS1 gene of SY_99 exhibits 98%,99% and 99% nucleotied sequence identity and 97%,99% and 99%amino acid sequence identity with other PPV strains NADL2_1,NADL2_2 and Kresse,respectively.Molecular cloning of SY_99 NS1 gene makes it possible to disclose the function of NS1 in PPV replication and prepare the diagnostic antigen with expressed NS1 protein.

关 键 词：猪 细小病毒 NS1基因 

分 类 号：S852.659.2[农业科学—基础兽医学] Q785[农业科学—兽医学]