研究在已有的新城疫病毒LaSota株反向遗传系统的基础上,利用新城疫病毒LaSota株作为表达载体,构建表达牛仪干扰素(bovine interferon α)完整开放阅读框的全基因组质粒pFL—BoIFNα,转染BHK-21细胞获得表达牛α干扰素重组新城疫病毒。采用RT—PCR方法检测,证实收获的鸡胚尿囊液内的重组病毒含有相应外源基因。将重组病毒尿囊液经紫外线照射灭活新城疫病毒,利用表达绿色荧光蛋白的重组水泡性口炎病毒(VSV—EGFP)在牛肾细胞(MDBK)上测定rL—BoIFNα抗病毒活性,证实表达牛干扰素的重组新城疫病毒尿囊液能有效抑制VSV—EGFP在牛肾细胞上的复制,rL—BoIFNα接种SPF鸡胚所收尿囊液抗病毒活性高达2×10^7U/mL。结果表明:牛α干扰素在重组新城疫病毒rL—BolFNα接种的SPF鸡胚尿囊液中获得良好表达,并具有高效而稳定的抗病毒活性。
The reverse genetic system of NDV La Sota strain have been established previously. In this study, a recombinant NDV containing bovine IFNα ( rL - BoIFNα) was constructed by inserting bovine IFNα into La Sota backbone. The results demonstrated that SPF embryonated egg infected with this virus express high levels of biologically active IFN. The allantoic fluid was exposured under UV rays to destroy infectivity of recombinant the allantoic fluid can reduced the replication of VSV - EGFP in cell culture. The results indicate type I bovine IFN (BoIFNα) has antiviral activity up to 2 × 10^7 IU/mL against VSV - EGFP in MDBK cell culture. The results indicate that the BoIFNα can be expressed in higher levels and have stable and high efficiency antiviral activity in vitro.