摘要
全长cDNA文库构建是改良胡椒性状的分子机理研究基础,有助于重要相关基因的克隆、功能分析、调控及其利用。以性状优良并具代表性的海南蒟（Piper hainanense）为试材,采用优化的In-Fusion SMARTerTMDirectional cDNA Library Construction Kit技术,构建叶片材料的全长cDNA文库。结果表明：文库的滴度为2.0×10^6cfu/mL,文库容量为1.3×10^6cfu。从文库中随机挑取20个克隆进行质粒DNA提取、PCR及酶切鉴定,结果显示：插入片段大小为1.0-2.0 kb,重组率为100%,表明该文库质量较高,可满足后续研究需要。
Piper nigrum L.is an economically important crop in the world.The construction of the full-length cDNA library of the leaf is the importantly fundamental work for the study on the molecular mechanism of P.nigrum L.,in which it can facilitate the cloning,characterization and utilization of the important genes involved.In this research,a full-length cDNA library of the leaf of P.hainanense,a well known piper cultivar in its high popularity in cultivation,was constructed with In-Fusion SMARTerTM Directional cDNA Library Construction Kit.The library has a capacity of 1.3× 10^6 clones,and its recombination efficiency was about 100%,and PCR results showed that the inserts sizes varied from 1.0 to 2.0 kb,suggesting that the cDNA library of P.hainanense was successfully constructed for the use in further study.
出处
《热带作物学报》
CSCD
北大核心
2013年第4期636-640,共5页
Chinese Journal of Tropical Crops
基金
外专局引智项目(No.20124600010)
热带香料饮料种质资源圃(No.12R22-20)
关键词
海南蒟
叶片
CDNA文库
构建
Piper hainanense
Leaf
CDNA library
Construction
