摘要
将鸡IFN-α和IFN-γ原核表达产物经Ni2+-NTA亲和层析法纯化,进行SDS-PAGE分析,再将纯化并已测定浓度的鸡重组IFN-α和IFN-γ稀释到1/10倍;用细胞病变抑制法检测重组表达蛋白的抗新城疫病毒活性,并利用感染病毒的鸡胚来测定鸡重组表达蛋白的抗病毒活性。结果显示,SDS-PAGE可检测到相对分子质量为25kU左右的蛋白条带,纯化的鸡IFN-α和IFN-γ抗新城疫病毒复制的活性分别为4.32×103 U/mg和2.65×103 U/mg;当鸡IFN-α和IFN-γ的比例为125∶1时,其联合抗新城疫病毒复制的活性最佳。抗鸡胚感染病毒活性的测定结果表明,与病毒对照组相比,IFN-α和IFN-γ及IFN-α和γ联合组,HA滴度都有明显的降低;并且从鸡胚的平均死亡时间上,联合干扰素对新城疫病毒的抑制作用更强于单个干扰素的应用。这证明了鸡IFN-α、IFN-γ及其联合干扰素对新城疫病毒的复制有明显的抑制作用。
Chicken IFN-α and IFN-γ were purified by Ni2＋-NTA affinity chromatography,SDS-PAGE analysis,then both chicken recombinant IFN-α and IFN-γ protein were diluted to 1/10-fold.Antiviral effect of the recombinant expression product was tested by cytopathogenic effect inhibition assay,and antiviral activity of chicken recombinant interferon were detected by using the infected chicken embryos.The results showed that about 25 kU protein bands can be detected by SDS-PAGE,the values of antiviral activity against NDV in BHK cell culture groups treated with Chicken IFN-α and IFN-γ were 4.32×103 U/mg and 2.65×103 U/mg respectively;The inhibitory activity against Newcastle disease virus replication showed the highest value occurred in the group that received the combination of recombinant interferon（IFN-α∶IFN-γ= 125∶1）.The results of antiviral activity on chicken embryos showed that the HA titers of IFN-α,IFN-γ and combination group were significantly lower than that of the virus control group.In terms of the average death time of chicken embryos,antiviral function of combination group was better than that of each IFN group.This proved that chicken IFN-α,IFN-γ and combination of interferon could significantly inhibit the replication of Newcastle disease virus.
出处
《上海交通大学学报（农业科学版）》
2011年第6期6-10,15,共6页
Journal of Shanghai Jiaotong University(Agricultural Science)
