以长白山区野生膜荚黄芪根和叶为试材,参照TRIzoL说明书提取总RNA,采用SMART技术构建其全长cDNA文库,为分子水平上揭示膜荚黄芪次生代谢调控机制提供充足的基因资源。结果表明:原始文库滴度为4.8×105 cfu/mL,重组率接近99.1%,插入片段大于0.5kb,平均大小1.54kb。所构建的cDNA文库的库容量、克隆效率和全长率满足挖掘膜荚黄芪次生代谢相关基因的要求。
Taking the roots and leaves of A.membranaceus of Changbai mountain as materials,total RNA was extracted reference the TRIzoL instructions,and the full-length cDNA library was constructed by SMART technology,in order to provided adequate gene resources for revealing the regulatory mechanism of secondary metabolism of Astragalus membranaceus at the molecular level.The results showed that the titer of unamplified library was 4.8×105 cfu/mL,the positive recombination rate was nearly 99.1%,the length of inserted fragments was larger than 0.5kb,and the average length was 1.54 kb.The storage capacity,recombination rate and full-length efficiency of the constructed library could meet the requirements of screening genes for secondary metabolism in A.membranaceus.