硬叶兜兰花芽SSH文库的构建

以硬叶兜兰花芽cDNA为Tester，营养芽cDNA为Driver，利用SMART策略构建了硬叶兜兰花芽的抑制性消减杂交(SSH)文库.通过PCR对文库中插入的片段进行检测后，筛选了288个插入片段为500bp以上的克隆进行测序.测序结果去除载体序列后聚类得到18条差异表达片段，用BLAST进行比对分析表明，这些差异表达基因所编码的蛋白与光合作用、合成代谢、基因调控等功能有关，其中包括多个转座子和反转录转座子的同源基因.

Construction of Suppression Subtractive Hybridization Library from the Floral Buds of Paphiopedilum Micranthum

With high ornamental and research value,Paphiopedilum micranthum is a species of the genus Paphiopedilum native to China,which is endangered and protected intensively by the country.However,there are few reports on its floral development mechanism because of the supply shortage and backward cultivation technology.In this study,a suppression subtractive hybridization(SSH) library was constructed using the cDNAs from the floral and vegetative buds of P.micranthum as the tester and the driver,respectively.The insert fragments were tested by PCR,and 288 clones with 500 bp or longer from the library were selected for sequencing.Totally,18 ESTs were obtained,after vector sequences removed and clustered.The ESTs were functionally annotated by Blast.Those which matched significantly with Nr database were further classified into functional groups including photosynthesis,biosynthetic metabolism,gene regulation,etc.Among them,a few sequences with homology to transposons or retro-transposons were obtained.The genes differentially expressed in floral buds of P.micranthum isolated in this study could make a base for investigating the molecular mechanism of floral development.