Gupta Dipinte, Ranjan Rajiv
Plant Biotechnology Lab, Department of Botany, Faculty of Science, Dayalbagh Educational Institute, Dayalbagh, Agra, 282005 India.
Virusdisease. 2020 Sep;31(3):369-373. doi: 10.1007/s13337-020-00617-8. Epub 2020 Aug 19.
CaMV35S is the most extensively used promoter for ectopic gene expression in plant system. However, multiple use of this promoter possesses several limitation i.e. homologous based gene silencing and differential suitability in monocot and dicot plants. The strength of a promoter is defined by the presence of -acting elements and trans acting nucleic binding factors, thus its strength can be regulated by changing the architecture of these regulatory elements. In the present study, eight hybrid promoters were designed from two parareteroviruses, rice tungro bacilliform viruses (RTBV) and mirabilis mosaic virus (MMV). The eight hybrid promoters, along with parental promoters were characterized for the presence of functional -elements and transcription factor binding sites (TFBS), which were predicted using bioinformatics tools such as PLACE and Matinspector. Presence of mirabilis mosaic virus modules for specific functions and over-represented modules was determined using Model inspector. A broad range of -elements (85), TFBS (1471) was obtained. Presence of Dehydration responsive element binding factors, Apetala 2 (AP2), WRKY, DNA binding with one finger DOF (DOFF) motifs had shown the functional relevance of these designed promoters with abiotic stress inducibility. In addition to these stress regulating TFBS, the presence of some enhancer like motifs such as P$OCSE, P$TERE, P$TODS, P$ASRC had shown the functional relevance of these promoters as a strong candidate for enhanced expression of ectopic gene.
花椰菜花叶病毒35S启动子(CaMV35S)是植物系统中异位基因表达最广泛使用的启动子。然而,多次使用该启动子存在一些局限性,即基于同源性的基因沉默以及在单子叶植物和双子叶植物中的适用性差异。启动子的强度由顺式作用元件和反式作用核酸结合因子的存在决定,因此其强度可以通过改变这些调控元件的结构来调节。在本研究中,从两种副杆状病毒,即水稻东格鲁杆状病毒(RTBV)和奇异果花叶病毒(MMV)设计了八个杂交启动子。这八个杂交启动子以及亲本启动子针对功能性顺式元件和转录因子结合位点(TFBS)的存在进行了表征,这些是使用诸如PLACE和Matinspector等生物信息学工具预测的。使用模型检查器确定了奇异果花叶病毒特定功能模块和过度表达模块的存在。获得了广泛的顺式元件(85个)、TFBS(1471个)。脱水响应元件结合因子、花器官特征基因2(AP2)、WRKY、单指DNA结合(DOFF)基序的存在表明了这些设计的启动子与非生物胁迫诱导性的功能相关性。除了这些胁迫调节TFBS外,一些增强子样基序如P$OCSE、P$TERE、P$TODS、P$ASRC的存在表明这些启动子作为异位基因增强表达的有力候选者的功能相关性。