西瓜果实特异性基因wml1的5’端上游调控序列的分离

ISOLATION OF 5' FLANKING REGION OF THE FRUIT-SPECIFIC GENE WML1 FROM WATERMELON BY UNEVEN PCR

西瓜(Citrullus vulgris Schrad.)ADP-葡萄糖焦磷酸化酶大亚基基因wml1的表达具有果实特异性。本文利用Uneven PCR技术成功地从西瓜基因组DNA中分离出一段长1864bp、位于wml1基因5’端上游的新序列。该序列含有TATA盒和CAAT盒,具典型的启动子特征。克隆序列中180bp-1752bp和958bp-1752bp两个片段分别与GUS基因融合进行瞬间表达试验,结果初步表明180bp-1752bp片段具有果实特异性启动子活性,转录调控元件位于序列的180bp-958bp。

The 5 flanking region of ADP-Glucose Pyrophosphorylase large subunit gene wmll, which covers 1864 bp, was isolated from watermelon genomic DNA by Uneven PCR. Sequence analysis indicated that there were putative TATA box and CAAT box at position 1237bp and 1284bp of the cloned fragment, respectively. Two transcriptional fusions between 5 regulational sequence of wmll and β-glucuronidase gene were constructed. Constructs containing 180bp-1752bp fragment could drive GUS gene transient express specifically in watermelon fruit, while 958bp-1752bp had no promoter activity. Putative fruit-specific cis-acting elements may locate between 180bp and 958bp.