CAJ | 학술논문

为进一步研究高致病性猪繁殖与呼吸综合征病毒（PRRSV）N 蛋白的结构、功能及免疫学特性，提取 PRRSV 云南分离株 A06 RNA，设计特异性引物扩增全长 N 基因，得到长度为372 bp 的目的片段，克隆至杆状病毒 Bac-To-Bac 表达载体 pFastBac HTB 中，转化大肠埃希菌 DH10Bac 进行同源重组，成功构建了含有 N 基因的真核表达载体，转染 Sf9细胞，回收得到重组杆状病毒。SDS-PAGE 和 Western blot 检测结果表明，表达产物17 ku，与预期大小一致，能与 PRRSV 阳性血清产生特异性反应。
위진일보연구고치병성저번식여호흡종합정병독（PRRSV）N 단백적결구、공능급면역학특성，제취 PRRSV 운남분리주 A06 RNA，설계특이성인물확증전장 N 기인，득도장도위372 bp 적목적편단，극륭지간상병독 Bac-To-Bac 표체재체 pFastBac HTB 중，전화대장애희균 DH10Bac 진행동원중조，성공구건료함유 N 기인적진핵표체재체，전염 Sf9세포，회수득도중조간상병독。SDS-PAGE 화 Western blot 검측결과표명，표체산물17 ku，여예기대소일치，능여 PRRSV 양성혈청산생특이성반응。
In order to study the N protein structure,function and immunological characteristics of porcine reproductive and respiratory syndrome virus,the A06 RNA was extracted from PRRSV of Yunnan strain. The specific primers were designed to amplify the full-length N gene,372 bp fragment was cloned into ex-pression vector pFastBac HTB of baculovirus Bac-To-Bac,and further homologous recombination in E.coli DH10Bac.The eukaryotic expression vector containing N gene was successfully constructed.Transfection of Sf9 cells,the recovery of recombinant baculovirus were made.SDS-PAGE and Western blot assay indi-cated that recombinant N protein was expressed successfully with molecular weight of 17 ku which was consistent with the expected size and could specifically react with anti PRRSV positive sera.