de Souza C R, Aragão F J, Moreira E C O, Costa C N M, Nascimento S B, Carvalho L J
Laboratório de Biologia Molecular, Universidade Federal do Pará, Belém, PA, Brazil.
Genet Mol Res. 2009 Mar 24;8(1):334-44. doi: 10.4238/vol8-1gmr560.
Cassava is one of the most important tropical food crops for more than 600 million people worldwide. Transgenic technologies can be useful for increasing its nutritional value and its resistance to viral diseases and insect pests. However, tissue-specific promoters that guarantee correct expression of transgenes would be necessary. We used inverse polymerase chain reaction to isolate a promoter sequence of the Mec1 gene coding for Pt2L4, a glutamic acid-rich protein differentially expressed in cassava storage roots. In silico analysis revealed putative cis-acting regulatory elements within this promoter sequence, including root-specific elements that may be required for its expression in vascular tissues. Transient expression experiments showed that the Mec1 promoter is functional, since this sequence was able to drive GUS expression in bean embryonic axes. Results from our computational analysis can serve as a guide for functional experiments to identify regions with tissue-specific Mec1 promoter activity. The DNA sequence that we identified is a new promoter that could be a candidate for genetic engineering of cassava roots.
木薯是全球6亿多人最重要的热带粮食作物之一。转基因技术有助于提高其营养价值以及对病毒病和害虫的抗性。然而,保证转基因正确表达的组织特异性启动子是必要的。我们利用反向聚合酶链反应分离了编码Pt2L4的Mec1基因的启动子序列,Pt2L4是一种在木薯贮藏根中差异表达的富含谷氨酸的蛋白质。生物信息学分析揭示了该启动子序列内的假定顺式作用调控元件,包括其在维管组织中表达可能所需的根特异性元件。瞬时表达实验表明Mec1启动子具有功能,因为该序列能够驱动GUS在菜豆胚轴中表达。我们的计算分析结果可为功能实验提供指导,以鉴定具有组织特异性Mec1启动子活性的区域。我们鉴定的DNA序列是一种新的启动子,可作为木薯根基因工程的候选者。