单子叶植物表达载体pUN3300的构建

植物双元表达载体在植物基因工程研究中具有重要作用,表达载体所包含的结构元件,如启动子、多克隆位点、筛选标记等,对植物遗传转化效率、外源基因表达强度及遗传稳定性等具有重要影响。本研究中,为提高目的基因对单子叶植物的遗传转化效率,对植物表达载体pCAMBIA3300进行改造,在原有以bar基因作为选择标记的基础上,采用不完全酶切的方法添加了Ubiquitin启动子,经酶切、测序表明,植物双元表达载体pUN3300构建成功。该载体对于研究外源基因功能、植物性状改良及新品种的培育,具有重要的价值。

Construction of Monocotyledon Expression Vector pUN3300

Plant binary expression vector has an important role in plant genetic engineering research.The structural elements which are included in the expression vector,such as promoter,multiple cloning sites,selection marker,have important influences on the efficiency of plant genetic transformation,intensity and stability of exogenous gene expression.In order to improve the efficiency of exogenous gene to target monocotyledon by genetic transformation,the commercialized expression vector pCAMBIA3300 was used and modified in this study.Using incompletely digestion technology,the ubiquitin promoter was successfully harvested and subcloned into pCAMBIA3300.The new constructed plant binary expression vector was named as pUN3300.The new vector of pUN3300 will be in favor of monocotyledon transformation and laid basis for the function analysis of exogenous genes,improvement of plant properties,and new variety breeding.