油菜APETALA3基因5′调控序列的克隆及分析

Isolation and Sequence Analysis of 5′ Regulatory Region of APETALA3 Gene from Brassica napus

利用单引物PCR方法获得了甘蓝型油菜APETALA3重复基因拷贝BnAP3-3和BnAP3-4的5′调控序列。分析结果表明,两段调控区序列一致性为80.43%,大多数的保守功能元件如TATA盒和CAAT盒没有明显差异,都含有一些应对环境胁迫及诱导的元件,如MYB1AT、GATABOX和DOFCOREZM等。少数顺式作用元件存在较大差异,其中CGACGOSAMY3、CArG box、CARGCW8GAT及CAREOSREP1为BnAP3-3的5′调控序列所特有,而BnAP3-4的5′调控序列则含有GT1CONSENSUS。半定量RT-PCR显示,BnAP3-3表达量远高于BnAP3-4,可能与二者调控区某些功能元件序列差异有关。

The 5′ regulatory region of two APETALA3 duplicated genes BnAP3-3 and BnAP3-4 in Brassica napus were cloned using single primer PCR.Sequence analysis showed that the two 5′ regulatory region sequences have 80.43% homologous with each other.Most of the conserved cis-regulatory elements were similar,such as TATA-box,CAAT-box and some stress responsive elements including MYB1AT,GATABOX and DOFCOREZM motifs.There were also a few different motifs between them.CGACGOSAMY3,CArG box,CARGCW8GAT and CAREOSREP1 motifs were found in the 5′ regulatory region of BnAP3-3,while the 5′ regulatory region of BnAP3-4 contained GT1CONSENSUS motif.The semi-quantitative RT-PCR demonstrated that the relative expression of BnAP3-3 was significantly higher than that of BnAP3-4.The results indicated that the cause of the expression difference of the two genes may be from the sequence difference of their 5′ regulatory region.