橡胶树HbFCA启动子在转基因烟草中的功能缺失分析

摘要

Using the 5' end serial deletion analysis method, five HbFCA promoter deletion fragments which had different distances from transcription start site were amplified by PCR, and they were used to replace the CaMV35S promoter of PBI121 carrier to obtain 5 HbFCA 5'deletion expression vectors, i.e. pA, pB, pC, pD and pE. The expression vectors were transformed into tobacco by using agrobacterium-meditated leaf disc method, and the well-rooted transformed tobacco plants in tissue culture vessels were directly detected with GUS staining. The results showed that the pE 267 bp, the shortest among the fragments, had promoter activity. The transient expression analysis of somatic embryo of rubber tree showed that the fragment pE also had better GUS staining, and could function as a HbFCA promoter.%利用5'缺失设计一系列引物,扩增H b FCA启动子质粒,获得5个启动子缺失片段,用这些片段分别替换PBI121载体中的CaMV35S启动子,得到5个HbFCA的5'缺失表达载体,即pA、pB、pC、pD和pE.利用农杆菌介导叶盘法进行烟草遗传转化,对组培瓶内生根较好的烟草叶片GUS染色,结果发现,长度为最小片段的276 bp的pE可以较好地实现HbFCA启动子的功能.对橡胶树体细胞胚瞬时表达检测发现,pE片段也能启动表达,有较好的GUS活性,可以实现HbFCA启动子的功能.