CAJ | 학술논문

根据鸭新城疫病毒 NP 基因序列设计1对特异性引物，RT-PCR 方法扩增出 NP 基因，克隆入原核表达载体 pET-30a，转化大肠杆菌感受态细胞 BL21（DE3），在 IPTG 的诱导下融合蛋白获得了成功表达，SDS-PAGE 结果显示表达的融合蛋白分子量约为59 kD，Western blotting 分析表明该重组融合蛋白能与 His 组氨酸单抗发生特异性反应。将目的蛋白切胶免疫 ICR 小鼠，制备了针对重组蛋白的多抗血清，Western-blotting 分析显示制备的多抗血清能够与鸭新城疫病毒感染的 SPF 鸡胚尿囊液总蛋白发生特异反应。以上结果表明，NP 基因在大肠杆菌中获得了成功表达，且制备的多抗血清可以用于 NP 蛋白的检测。
근거압신성역병독 NP 기인서렬설계1대특이성인물，RT-PCR 방법확증출 NP 기인，극륭입원핵표체재체 pET-30a，전화대장간균감수태세포 BL21（DE3），재 IPTG 적유도하융합단백획득료성공표체，SDS-PAGE 결과현시표체적융합단백분자량약위59 kD，Western blotting 분석표명해중조융합단백능여 His 조안산단항발생특이성반응。장목적단백절효면역 ICR 소서，제비료침대중조단백적다항혈청，Western-blotting 분석현시제비적다항혈청능구여압신성역병독감염적 SPF 계배뇨낭액총단백발생특이반응。이상결과표명，NP 기인재대장간균중획득료성공표체，차제비적다항혈청가이용우 NP 단백적검측。
According to the genome sequences of the duck NDV,a pair of specific primers was designed.The NP gene of the virus was amplified by RT-PCR and cloned into the prokaryotic expression vector pET-30a.The recombinant plasmids were transformed into BL21 (DE3)E.coli.Subsequently,this recombinant fusion protein was successfully expressed following IPTG induction.SDS-PAGE showed that the protein had an approximate molecular weight of 59 kD,and the Western blotting assay revealed that it could be recognized by the monoclonal antibody against histidine-tagged protein.The antiserum was,then,produced by the immunized ICR mice with the recombinant protein.Western blotting on the antiserum indicated its specific reactivity with NDV allantoic fluid.It was concluded that theNP gene could be successfully expressed in E.coli,and the antiserum could be used for detection of NP protein.