Cloning and Functional Analysis of a Promoter of ?-conglycinin Subunit Gene in Soybean / 中国生物工程杂志

ABSTRACT

A promoter fragment (7S?P ) of ? 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 7S?P fragment from these five soybean accessions shared 99% homology, this indicated that the promoter regions of ? subunit gene were conserved. Meanwhile , sequencing analysis showed that the 7S?P fragment contained several seed-specific motifs, such as RY motif, AGCCCA motif, ACGT motif and A/ T rich motif. The expression vector pBI121-7S?P was constructed with the 7S?P 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 7S?P 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 7S?P promoter. The results showed that 7S?P was a seed-specific promoter.