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通过对模板DNA、Primer、dNTPs、10×Taq 10 Buffer、Taq酶的不同浓度进行单因素筛选,并对退火温度和循环次数进行摸索,建立和优化了灰葡萄孢菌Flipper转座子PCR扩增体系。采用该体系及Flipper转座子引物扩增得到了1 159 bp的片段,通过比对发现,该片段与Botryotinia fuckeliana的Flipper转座因子转座酶基因相似性达99%,表明扩增得到的是预期目的片段。Flipper转座子扩增体系的建立,为深入研究我国葡萄灰霉病菌菌株的转座子类型及其与致病力、抗药性等因素之间的关系奠定了重要的基础。
Through single-factor screening of different concentrations of template DNA, Primer, dNTPs, 10 × Taq 10 Buffer and Taq DNA polymerase, the annealing temperature and the number of cycles were explored to establish and optimize the PCR amplification of Flipper transposon from Botrytis cinerea system. A fragment of 1 159 bp was obtained by using this system and Flipper transposon primer. The similarity of Flipper transposase transcriptase gene of Botryotinia fuckeliana was 99%, which indicated that the amplified fragment was Expected purpose fragment. The establishment of Flipper transposon amplification system lays an important foundation for further study on the transposon type and its relationship with pathogenicity, drug resistance and other factors in our country.