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对抗病的朝天椒地方品种,以紫外线(UV)处理叶片并提取叶片总RNA,采用SMARTTMcDNA文库构建试剂盒构建了一个cDNA文库.该文库含有2.6×106个独立的噬菌斑克隆,扩增后滴度达到1.5×109pfu.μL-1,随机挑取的噬菌斑PCR检测发现重组率达90.0%.在搜索、拼接与植物WRKY、bZIP、AP2和E2等重要调节基因同源的辣椒表达序列标签(ESTs),获得重叠群的基础上,分别设计上述基因ESTs重叠群特异性引物,以cDNA文库为模板进行PCR扩增.结果表明,部分上述基因cDNA存在于文库中.因此,本研究所构建的文库可用于进一步分离响应UV的重要调节基因cDNA的分离.
For the local cultivated Asparagus officinalis, a cDNA library was constructed by using SMARTTM cDNA library to construct a cDNA library by treating the leaves with UV light and extracting the total RNA of the leaves. The library contains 2.6 × 106 independent plaque clones, After the titers reached 1.5 × 109 pfu.μL-1, the PCR results showed that the recombination rate was 90.0% .Sequencing and searching for the expression of pepper genes homologous to important regulatory genes WRKY, bZIP, AP2 and E2 Sequence tags (ESTs) to obtain contigs, we designed the specific primers of contigs ESTs of these ESTs respectively, and amplified them by PCR using the cDNA library as a template.The results showed that some of the above cDNAs existed in the library.Therefore, The constructed library can be used for further isolation of cDNAs that are important regulators of UV response.