拟南芥中花特异表达启动子PCHS的分离及其功能初探

Cloning and Function Analysis of a Flower-specific Expression Promoter PCHS

根据已发表的拟南芥启动子序列(AF248988)设计合成一对引物,以拟南芥(Arabidopsis)基因组DNA为模板,通过PCR技术获得了约含500bp大小的DNA片段,经纯化后测序得到531bp的目的片段。通过DNAman进行序列分析表明,与文献报道的PCHS启动子序列有99%的同源,含有4个花特异表达的调控元件。将此启动子替换pBI121上的CaMV35S启动子构建植物表达载体,农杆菌介导法浸染矮牵牛花、茎、叶、根。GUS瞬时表达结果表明,在花上出现较深的蓝色,而在茎上的蓝色很浅,在根和叶中不显蓝色,初步确定该启动子具有较强的花特异表达特性。

A polymerase chain reaction（PCR）amplification was used to isolate a flower-specific expression promoter PCHS from Arabidopsis genomic DNA. Sequence analysis revealed that PCHS was 531 bp in total length and shared significant nucleotides identity 99% with GenBank databases, which contained four flower-specific expression regulatory elements. Intermediate vector pPCHS was constructed by inserting the promoter to the upstream of GUS in pB1121, where the original CaMV35S promoter was replaced by the PCHS promoter. The flower, stem, leaf ,and root of petunia hybrida were transformed via Agrobacterium tumefaciens. Histochemical localization of GUS activity indicated that the PCHS promoter could confer a flower-specific expression pattern to GUS.