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发明公开
CN110343671A 一种表达VP2基因的重组Ⅰ型马立克氏病病毒疫苗毒株 审中-实审

一种表达VP2基因的重组Ⅰ型马立克氏病病毒疫苗毒株
摘要：
本发明构建了一株表达传染性法氏囊病毒VP2基因的重组鸡马立克氏病病毒疫苗株并对其应用进行了探究。本发明利用同源重组技术，将包含传染性法氏囊病毒VP2基因和CMV启动子序列的基因片段CMV-VP2及含flp识别位点的Kana基因插入到鸡马立克氏病病毒弱毒疫苗SC9-1株的UL45/UL46基因之间，构建获得在UL45/UL46基因间插入CMV-VP2表达框和Kana表达框的重组质粒，而后通过阿拉伯糖将Kana基因诱导掉，得到只含有CMV-VP2表达框的重组质粒，由其拯救获得表达传染性法氏囊病毒VP2基因的重组鸡马立克氏病病毒疫苗株。研究表明，本发明获得的疫苗株具有与亲本毒SC9-1疫苗株同等的体外复制能力以及良好的遗传稳定性，并且能够同时抵抗MDV超强毒株以及IBDV超强毒株的攻击。由此可见，本发明获得的表达IBDV-VP2基因的重组MDV疫苗株可用于预防鸡传染性法氏囊病和马立克氏病。
摘要（英）：

The invention constructs a recombinant chicken Marek's disease virus vaccine strain expressing an infectious bursal disease virus VP2 gene and explores the application of the recombinant chicken Marek's disease virus vaccine strain. By utilizing a homologous recombination technology, a gene fragment CMV-VP2 containing an infectious bursal disease virus VP2 gene and a CMV promoter sequence and a Kana gene containing flp recognition locus are inserted between UL45/UL46 genes of a chicken Marek's disease virus attenuated vaccine SC9-1 strain to construct and obtain a recombinant plasmid with a CMV-VP2 expression cassette and a Kana expression cassette inserted between the L45/UL46 genes, then the Kana gene is induced off by arabinose to obtain a recombinant plasmid only containing the CMV-VP2expression cassette, and the recombinant chicken Marek's disease virus vaccine strain expressing the infectious bursal disease virus VP2 gene is obtained by its rescue. Studies show that the obtainedvaccine strain has the same in-vitro replication capacity and good genetic stability with a parent virus SC9-1 vaccine strain, and can simultaneously resist the attack of an MDV super-virulent strainand an IBDV super-virulent strain. Thus, it can be seen that the recombinant MDV vaccine strain expressing the IBDV-VP2 gene can be used for preventing an infectious bursal disease and a Marek's disease of chickens.

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