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低毒病毒/板栗疫病菌是一个研究病毒宿主相互作用的优良的实验系统。为了更好的了解病毒与宿主相互作用的机制,我们建立了一个野生型无病毒板栗疫病菌菌株EP 155和受低毒病毒感染的菌株EP 713的cDNA混和文库,以进行表达序列标签(EST)测定。对543个克隆的外源片段的分析表明,该文库克隆中含外源片段的比例为89.8%;通过小规模测序,发现空质粒与小片段克隆占测序总数的10.1%;表达丰度最高的4个克隆出现次数的总和占测序总数的9.6%。通过设计引物进行PCR和原位杂交,对cDNA文库2万多个克隆进行了筛选,共筛除了占总数19.7%的空质粒与小克隆和5.5%的高丰度克隆,为高效率的大规模EST测序准备了条件。
The virulent virulent / chestnut blight bacteria is an excellent experimental system for studying viral host interactions. In order to better understand the mechanism of virus-host interaction, we constructed a cDNA hybrid library of the wild-type virus-free T. chestnut strain EP 155 and the low-virus-infected strain EP 713 for expression of sequence tags (ESTs) Determination. Analysis of exogenous fragments of 543 clones showed that the percentage of exogenous fragments contained in this library clone was 89.8%. By small-scale sequencing, it was found that the empty plasmid and small fragment clone accounted for 10.1% of the total number of sequencing; the highest expression abundance The sum of the number of 4 clones accounted for 9.6% of the total number of sequencing. By designing primers for PCR and in situ hybridization, more than 20,000 clones of the cDNA library were screened, screened for 19.7% of the total number of empty plasmids and small clones and 5.5% of high abundance clones, for a high-efficiency large-scale EST sequencing prepared conditions.