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为理解荷花Nelumbo nucifera花器官转录组表达情况,分别选取不同花型的代表品种‘洪湖红莲’Nelumbo nucifera‘Honghu Honglian’(单瓣)、‘唐招提寺莲’N.nucifera‘Tangzhaotisi Lian’(重瓣)和‘千瓣莲’N.nucifera‘Qianban Lian’(千瓣及全重瓣)的花蕾为材料分离mRNA,利用SMART技术合成双链cDNA,经限制性内切酶Sfi I酶切后回收去掉接头和500 bp以下片段的c DNA。将c DNA与p UC19载体连接,构建荷花花蕾cDNA文库。经检测,该文库容量为1.12×106 pfu·m L-1,插入片段大小集中在500~2000 bp,重组率为95%。该文库的成功构建为荷花花蕾期转录组数据的开发及其花器官发育相关基因的功能研究奠定了分子基础。
In order to understand the expression of organ transcriptome of Nelumbo nucifera, the representative varieties of Nelumbo nucifera’Honghu Honglian ’,’ Nnucifera’Tangzhaotisi Lian ’ Double-stranded cDNA was synthesized by SMART technique and digested with restriction endonuclease Sfi I Recover the c DNA from the adapter and fragments below 500 bp. The c DNA was ligated with the pUC19 vector to construct a lotus flower bud cDNA library. After testing, the library size was 1.12 × 106 pfu · m L-1, the size of the insert was 500-2000 bp, and the recombination rate was 95%. The successful construction of this library lays the foundation for the development of flower bud stage transcriptome data and the function study of flower-related organ development-related genes.