Abstract A promoter fragment (7SαP ) of α subunit gene was isolated by PCR from genomic DNA in various soybean accessions, including cultivars Nannong99-10,N2899,Nannong 88-1, and wild soybeans Jiangpu -1 and ZYD4174.The sequences of 7SαP fragment from these five soybean accessions shared 99% homology, this indicated that the promoter regions of α subunit gene were conserved. Meanwhile , sequencing analysis showed that the 7SαP fragment contained several seed-specific motifs, such as RY motif, AGCCCA motif, ACGT motif and A/ T rich motif. The expression vector pBI121-7SαP was constructed with the 7SαP fragment (from Nannong99-10) and the GFP reporter gene for functional analysis. Arabidopsis plants were transformed by Agrobacterium mediated method. Southern blot results showed that the 7SαP had been integrated into the genome of Arabidopsis. Assay of GFP expression in the seeds of transgenic Arabidopsis was determined to identify the function of 7SαP promoter. The results showed that 7SαP was a seed-specific promoter.
|
