关键词: 甘菊, 甜菜碱醛脱氢酶基因, 启动子, 克隆, 瞬时表达

Abstract:

In order to provide inducible promoter for chrysanthemum [Dendronthema ×grandiflora (Ramat.) Kitam.] transgenic breeding, referring to the strategy of 5′RACE, four promoter sequences of betaine aldehyde dehydrogenase (BADH) gene from Dendranthema lavandulifolium (Fisch. ex Trautv.) Makino were cloned by anchored PCR walking, which were named DBP11, DBP12, DBP21 and DBP22 (GenBank accession No. DQ497620~DQ497623). The four sequences are 1 230 bp, 1 249 bp, 1 273 bp and 574 bp long respectively. The homology of the corresponding regions between every two sequences is above 89%. DBP12 and DBP21 are the promoters of DlBADH1 and DlBADH2 (GenBank accession No. DQ011151 and DQ011152), DBP11and DBP22 are the promoters of other members in BADH gene family from Dendranthema lavandulifolium. Many cis-acting elements related to water stress and ABA inducement were found in all the sequences. New expression vectors were constructed by replacing the 35S-CaMV promoter with the above promoter sequences to drive the reporter gene GUSplus of the expression vector pCAMBIA1305.2. The new vectors were transferred into Agrobacterium to infect leaf disks of Dendranthema lavandulifolium. The result of transient expression indicated that all the sequences had the function to drive reporter gene.

Key words: Dendranthema lavandulifolium, betaine aldehyde dehydrogenase (BADH) gene, promoter, cloning, transient expression