利用实时定量PCR方法,检测大豆酰基载体蛋白硫酯酶(acyl-ACP thioesterase)基因在大豆各组织中的表达方式,结果显示该基因在大豆根、茎、叶、花中的表达活性低,而种子中的表达活性较高。利用PCR方法,克隆大豆acyl-ACP thioesterase基因5'端上游2 057 bp序列,命名为AP。在线启动子预测软件分析结果表明AP序列中含有多种典型的种子特异表达元件,如RY repeat、SEF1 motif、SEF3 motif、SEF4 motif、E-box、ACGT等顺式作用元件,推测大豆acyl-ACP thioesterase基因启动子具有种子特异表达活性。
The expression pattern of soybean acyl-ACP thioesterase gene in soybean tissues was detected by real-time quantitative PCR (RTQ-PCR). The result showed that there were a little activity in roots, stems,leaves and flowers, but there was higher activity in seeds. The 5'-flanking upstream sequence of soybean acyl-ACP thioesterase gene, named AP, was isolated from soybean genomic DNA by PCR method, and the length was 2 057 bp. Sequence analysis by PLACE revealed that this fragment contained a series of motifs related to seed-specific promoters ,such as RY repeat, SEF1 motif, SEF3 motif, SEF4 motif, E-box and ACGT. It can be inferred that AP promoter possess the function driven downstream gene expression exclusively in soybean seeds.