【摘要】 参照NDVLa Sota株核酸序列(AF077761)设计1对引物,利用RT-PCR扩增F基因并得到了长为1 700 bp的片段,将其克隆到pGEM-T easy vector中,经酶切鉴定和测序后克隆进原核表达载体pET-32a,将重组表达质粒转化BL21(DE3),在IPTG诱导下表达约83 kd的融合蛋白,SDS-PAGE电泳和Western blotting检测证实该基因片段获得高效表达且表达产物具有免疫学活性。更多还原

【Abstract】 A pair of primers was designed according to the sequence of F gene of NDV La Sotastrain(AF077761) for amplifying the F gene by RT-PCR.1 700 bp DNA fragment was amplified and cloned into pGEM-T easy vector.After restriction endonuclease analysis and sequencing,the fragment was inserted into the expression vector pET-32a.The recombinant plasmid produced a 83 kd fusion protein in E.coli BL21(DE3) under the induction of IPTG.The expression quantity and immunoreactivity of the fusion protein was detected by SDS-PAGE electrophoresis and Western blotting.更多还原