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将黄瓜授粉前后多个发育阶段的幼果组织等量混合后提取总RNA和mRNA,以λTriplEx2为载体、XL1-Blue为宿主菌,构建了1个黄瓜幼果cDNA文库;其滴度为1.165×106pfu/mL,重组率在94.4%左右.测序获得116条EST,92.2%的长度在400bp以上,19%为重叠序列.在GenBank中进行BLAST分析后确认与已知功能基因相似的EST序列有71条,有相似序列而功能未知的基因和没有相似序列的EST序列各占19.83%和18.97%.从对文库的检验结果看,构建的cDNA文库重组率较高,库容达到预期要求.
The total RNA and mRNA were extracted after equaling the young fruit tissues of multiple developmental stages before and after the pollination of cucumber. Using λTriplEx2 as the vector and XL1-Blue as the host bacteria, a cucumber young fruit cDNA library was constructed with a titer of 1.165 × 106pfu / mL, the recombination rate was about 94.4%, 116 ESTs were sequenced, 92.2% of them were over 400bp in length and 19% were overlapped sequences.After BLAST analysis in GenBank, ESTs with similarities with known functional genes were identified as 71 19.83% and 18.97% of the ESTs with similar sequences and unknown function, respectively, and 18.97% of the EST sequences with no similar sequences.According to the test results of the library, the recombination rate of the constructed cDNA library is higher and the storage capacity meets the expected requirement.