[关键词]
[摘要]
目的 通过构建茉莉酸甲酯(MeJA)处理的白木香cDNA文库,为揭示伤害诱导沉香倍半萜合成的分子机制奠定基础。方法 以MeJA处理的白木香愈伤组织为材料,以改造的pGADT为载体,利用SMART技术构建白木香全长cDNA文库。结果 构建的文库原始滴度为8.99×106 pfu/mL,阳性克隆子的比例为97%,插入片段的大小在500~3 000 bp,平均大于1 000 bp。结论 构建的文库无论从库容量、克隆效率,还是全长率方面评价,均达到了高质量文库的要求。
[Key word]
[Abstract]
Objective To construct the cDNA library of Aquilaria sinensis treated with methyl jasmonate (MeJA), and to lay the foundation for revealing the molecular mechanism of damage-induced sesquiterpene synthesis of Aquilariae Lignum Resinatum. Methods The full-length cDNA library was constructed by SMART technology with the MeJA-treated A. sinensis calli as materials and the modified pGADT as vector. Results The original titer of the unamplified library was 8.99 × 106 pfu/mL, the percentage of the positive recombination was 97%, and the length of inserted fragments was 500—3 000 bp with the average length of more than 1 000 bp. Conclusion The constructed library in this study reaches the high-quality library requirements in terms of storage capacity, recombination rate, or full-length cloning efficiency.
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[基金项目]
国家自然科学基金资助项目(31100220,81173481)