The NS2 gene of Rice stripe virus(RSV) was amplified by RT-PCR,cloned into pGEM-T vector and sequenced.The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2.The ...The NS2 gene of Rice stripe virus(RSV) was amplified by RT-PCR,cloned into pGEM-T vector and sequenced.The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2.The recombinant plasmid was introduced into Escherichia coli strain BL21(DE3) pLysS.SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG.The recombinant NS2 protein was purified with Ni2+-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit.NS2 protein was successfully detected in small brown planthopper(Laodelphax striatellus) at 1∶1 600 dilution of the total protein of single planthopper and in infected rice(Oryza sativa) at 1∶800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody.展开更多
文摘The NS2 gene of Rice stripe virus(RSV) was amplified by RT-PCR,cloned into pGEM-T vector and sequenced.The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2.The recombinant plasmid was introduced into Escherichia coli strain BL21(DE3) pLysS.SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG.The recombinant NS2 protein was purified with Ni2+-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit.NS2 protein was successfully detected in small brown planthopper(Laodelphax striatellus) at 1∶1 600 dilution of the total protein of single planthopper and in infected rice(Oryza sativa) at 1∶800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody.
