【作者】 康宁；

【导师】 陈保善；

【作者基本信息】 广西大学 ， 生物化学与分子生物学， 2004， 硕士

【摘要】 病毒与宿主相互作用是现代病毒学研究最重要的前沿领域之一。本文采用具有独特优点的一个崭新的病毒／宿主模型系统—低毒病毒／板栗疫病菌系统来研究病毒与宿主的相互作用。建立了板栗疫病菌的原生质体再生脱毒方法，该方法可望应用到其他具有工农业重要使用价值的感染病毒真菌的脱毒纯化。通过用EP155单交换随机整合质粒文库转化低毒的板栗疫病菌工程菌株LC1，构建了突变体库，并以孢子形成为标记，从突变体库中筛选到了一批携带病毒且产孢良好的病毒靶基因突变株。通过对突变株B910 Southern杂交分析确定为单插入突变。质粒拯救获得了突变基因片段，对该基因片段序列分析表明，该基因(命名为cpta1)的编码产物与炭疽菌Colletotrichum lindmuthianum中的转录激活因子CLTA1有64％的同源性和75％的相似性。这些结果暗示CPTA1可能是低毒病毒的一个靶标，其功能因病毒侵染而发生改变并且干扰真菌的正常产孢过程。更多还原

【Abstract】 The interaction between virus and host is one of the most important front of virology. A novel hypovirus/Cryphonectria parasitica system was adopted to study the interaction between a virus and its host. It was found that the hypovirus could be eliminated after protoplast regeneration in C. parasitica, a method that can be used to eliminate virus for other fungi. By insertional mutagenesis of the transgenic strain LCI with a single-cross random integration plasmid library that contain random DNA of the virus-free wild type C. parasitica, many mutants with varied phenotype were obtained. One of such mutants, B910, sporulated well while was still infected by the viruses. Southern blot analysis showed it was a single insertional mutant. A fragment of 888bp that flanks the insertion site was recovered by plasmid rescue. Sequence analysis indicated that the insertion interrupted an ORF (termed CPTAl) that encodes a protein similar to a GAL4-like transcription activator of Colletotrichum lindmuthianum with 64% identity and 75% positives. This result implies that CPTAl maybe the target of the hypovirus and whose funtion was modulated by the virus to interfere with the sporulation process of the fungus.更多还原