摘要
利用根癌农杆菌介导的遗传转化,将启动子诱捕(Promotertrapping)元件插入到棉花基因组,获得141个独立的转化子,其中97%的转化子经PCR扩增为阳性。不同组织中GUS基因的表达频率为:根部48%,茎的微管组织9.2%,叶5.2%,花51%;同时检测了不同植株中GUS基因的表达模式,发现GUS基因在不同株系的植株间的表达模式呈现较大的差异,有些植株中GUS基因是组织特异表达,有些则是器官特异表达,有些则在多个器官中均有表达。所建立的启动子诱捕系统中的GUS基因高频率、多模式和时空特异性表达为分离基因及其调节序列、开展功能基因组研究奠定了坚实的基础。
The technique of promoter trapping was developed to investigate its viability in cotton （ Gossypium hirsutum L. ） functional genomics. 141 independent transformants of cotton were generated via Agrobacterium tumefaciens mediated transformation, of which 97% showed positive by PCR detection. The reporter, GUS gene,was expressed to different extent in different organs,with a frequency of 48% in roots,9.2% in vascular bundles of stem ,5.2% in leaves,and 51% in flowers. Meanwhile,we discovered that there existed great differ- ences in expression patterns among different transgenic lines. Their GUS expression patterns were organ- or tissue-specific or ubiquitous in all of the plants. The promoter trapping system developed here was characterized as an effective method for creating mutants with diverse reporter gene expression patterns, which laid a solid foundation for further research of functional genomics in cotton.
基金
国家自然科学基金项目(编号:30471109)~~
