CAJ | 학술논문

将新城疫病毒(newcastle disease virus,NDV)感染健康鸭胚,采用尿囊液方法提取总 RNA,根据 Gen-Bank已发表的新城疫病毒HN基因序列设计一对引物,以一株鸭源 NDV 的总 RNA 为模板,经 RT-PCR 扩增出约1.7 kb的HN基因片段.将其克隆到原核表达载体 pET-30a,获得重组质粒 pET-HN 后,转化大肠杆菌 BL21(DE3)感受态细胞.重组菌经 IPTG诱导表达后,裂解产物进行 SDS-PAGE分析,结果发现在约64 ku 处可见预期条带.Western-blot结果表明：表达的 HN蛋白可与 NDV阳性血清发生特异性结合,具有良好的反应原性,本结果为鸭源 NDV抗体检测提供了理论依据.
장신성역병독(newcastle disease virus,NDV)감염건강압배,채용뇨낭액방법제취총 RNA,근거 Gen-Bank이발표적신성역병독HN기인서렬설계일대인물,이일주압원 NDV 적총 RNA 위모판,경 RT-PCR 확증출약1.7 kb적HN기인편단.장기극륭도원핵표체재체 pET-30a,획득중조질립 pET-HN 후,전화대장간균 BL21(DE3)감수태세포.중조균경 IPTG유도표체후,렬해산물진행 SDS-PAGE분석,결과발현재약64 ku 처가견예기조대.Western-blot결과표명：표체적 HN단백가여 NDV양성혈청발생특이성결합,구유량호적반응원성,본결과위압원 NDV항체검측제공료이론의거.
Infecting healthy duck embryo with NDV,and the HN gene segments of 1 .7 kb from duck were amplified by RT-PCR,the primers were designed on the basis of HN gene sequence of NDV in Gen Bank.Then,the HN gene segments were inserted into the prokaryotic vector pET-30a and transformed in-to E.coli BL21(DE3).After that,the proteins were successfully expressed by IPTG induction,SDS-PAGE showed that the fusion protein had a molecular weight about 64 KDa.The result of Western-blot showed that HN protein could be recognized by serum against NDV.This study provided a certain value in duck-or-igin NDV antibody detection.