草豆蔻cDNA文库的构建及鉴定

Construction and identification of cDNA library of Alpinia hainanensis (Zingiberaceae)

以草豆蔻花序原基为材料,构建了cDNA文库。原始文库滴度为0.8×106pfu/mL,扩增后滴度为4.23×1011pfu/mL。插入片段大小在500bp～1.5kb之间,重组率为95.3%。以水稻RAP1A基因中包含MADS-box保守区段的序列为探针对该文库进行筛选,获得的阳性克隆经测序及序列比对分析,确认其中共有10个含MADS-box的阳性克隆。

An infloreseenees primordium eDNA library of Alpinia hainanensis K. Schumann was constructed. The titer of the eDNA primary library is 0. 8×10^6 pfu/mL,while that of the amplified library is 4. 23 ×10^11pfu/mL. The recombinant rate is about 95. 3 %, and the length of eDNA is between 500 bp- 1.5 kb. The fragment containing the MAD,S-box conserved sequence of RAP1A gene of Oryza sativa is used as probe to screen the library. Positive clones are sequenced and the sequences are analyzed first blast within the NCBI database, then alignment with the known MAD-box gene. Ten MADS-box genes from A. hainanensis are identified.