茉莉酸甲酯诱导的白木香cDNA文库的构建及初步鉴定  

Construction and primary authentication for cDNA library of Aquilaria sinensis incuced by methyl jasmonate

作　　者：徐艳红[1] 杨欣[1] 梁良[1] 高志晖[1] 张争[1,2] 魏建和[1,2] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]中国医学科学院北京协和医学院药用植物研究所海南分所,海南省南药资源保护与开发重点实验室,海南万宁571533

出　　处：《中草药》2014年第1期102-106,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(31100220,81173481)

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8．99×10^6pfu／mL，阳性克隆子的比例为97％，插入片段的大小在500-3000bp，平均大于1000bp。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。Objective To construct the cDNA library ofAquilaria 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.

关 键 词：白木香 茉莉酸甲酯 CDNA文库 愈伤组织 沉香 

分 类 号：R282.12[医药卫生—中药学]