利用抑制性差减杂交技术筛选草原龙胆花器官发育特性基因  

Primary Screening of Homeotic Genes During Floral Development with Suppression Subtractive Hybridization in Lisianthus (Eustoma grandiflorum)

作　　者：徐启江[1] 关录飞[1] 吴笑女[1] 李玉花[1] 

机构地区：[1]东北林业大学花卉生物工程研究所,黑龙江哈尔滨150040

出　　处：《生物技术通讯》2007年第3期401-404,共4页Letters in Biotechnology

摘　　要：目的:探讨草原龙胆花发育的分子机制,为进一步阐述花器官同源异型、属于MADS-box基因家族的一系列基因在调节开花植物花瓣和雄蕊的发育中的作用奠定基础。方法:以草原龙胆不同发育时期的花器官(萼片、花瓣、雄蕊、雌蕊)原基的cDNA作为试验方(tester),以茎叶组织的cDNA作为驱动方(driver),利用抑制性消减杂交技术构建了一个富集花器官发育特性基因的抑制性差减cDNA文库。对抑制性差减cDNA文库进行筛选、测序及Blast同源性比较。结果:获得了与花器官发育相关的特异性基因。结论:构建了抑制性差减cDNA文库,为克隆草原龙胆花器官发育特异性基因全长序列奠定了基础。Objective： To analyze the molecular mechanisms of floral development in lisianthus （Eustoma grand/florum）, above which paves the way to further elaborate how do the genes, which belong to the MADS-box gene family, played an important roles in regulating the development of floral organ development in flowering plants. Methods： Suppression subtractive hybridization technology was applied in the search of floral organ identity genes during floral development in lisianthus. A subtractive cDNA library, which was enriched for genes related to flower organ characteristics, was constructed. The cDNA prepared from flower organs of sepal, petal, androecium, and gynoecium at the different stage of development was used as the tester and the cDNA from the stem and leaf organ as the driver. Results： The identity genes expressed during floral development in lisianthus were searched. From the library, high quality sequences were obtained. By running blastn program of NCBI, the homologous genes with the network data base were obtained. Conclusion： The results showed that the subtractive cDNA library was constructed successful. The results establish the foundation for cloning the whole length of the homeotic genes during floral development.

关 键 词：草原龙胆 抑制性消减杂交 花器官特性基因 

分 类 号：Q78[生物学—分子生物学]