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为了对山东省滨州市周边地区6株新城疫流行情况进行研究,试验对不同毒株的治病力进行测定,采用RT-PCR技术扩增F基因,利用Meg Align软件进行序列分析,同时克隆DZNDV/007株F蛋白的两个抗原表位F72和F161,构建两个融合表达质粒p ET-32a-F72和p ET-32a-F161,并对其进行诱导表达及Western-blot分析。结果表明:5株属于基因Ⅱ型,为弱毒株;1株属于基因Ⅶ型,为强毒株;IPTG诱导得到两个大量表达的融合蛋白His-F72和His-F161,且具有良好的反应原性。说明新城疫基因Ⅶ型毒株已成为我国的主要致病毒株。
In order to study the epidemic situation of 6 Newcastle disease in the surrounding area of Binzhou, Shandong Province, the pathogenicity of different strains was tested. The F gene was amplified by RT-PCR and sequenced by Meg Align software. Meanwhile, DZNDV / 007 strain of F protein F72 and F161 antigen epitopes to construct two fusion expression plasmid p ET-32a-F72 and p ET-32a-F161, and its expression and Western-blot analysis. The results showed that five strains belonged to genotype Ⅱ and the most virulent strains. One strain belonged to genotype Ⅶ, which was a virulent strain. Two large fusion proteins, His-F72 and His-F161, were induced by IPTG. Sex. That Newcastle disease virus genotype Ⅶ has become China’s major virulent strains.