![]() ![]() atroviride, with the aim of confirming DAPG can be biosynthesized heterologously, in eukaryotic cells and in T. ![]() We designed three steps to prove that DAPG can be synthesised in T. We successfully cloned the phlABCD via colony PCR from Pseudomonas fluorescens 2P24. Further studies shows that phlABCD are the DAPG synthesis genes. And it has been demonstrated that a cluster named phl responsible for the biosynthesis of DAPG, which contains eight genes, from phlA to phlH. Īs has mentioned before, DAPG is initially found in specific strain of Pseudomonas fluorescens. Last but not least, previous studies comfirmed that DAPG almost has no harmful impact on human health under the effective concentration, and can be degraded easily in nature. atroviride, which to some extent reduces the cost. What’ s worth to mention is that the DAPG our device releasing can be synthesized by the engineered T. atroviride nearby to remove the phlF protein binding to the phlO domain so as to turn on the expression of corresponding gene that is responsible for reporting signals or inhibition of pathogens. The molecules diffused into our engineered T. ![]() When our device detected that the plant is infected with pathogens and is in poor condition by analyzing the changes in volatile organic compounds (VOC) given off by plants, it will release a few DAPG molecules as signal to communicate with T. We take advantage of DAPG-Off/On System to make our device interact with our engineered T. This year we aim to integrate both hardware and engineered organisms to provide a new blue print of automatic agricultural. What’s more, it is demonstrated that the transcriptional regulation is rapid, so this set of system has strong potential to be applied to several signal transduction pathways thanks to its supersensitive feature. DAPG remove the phlF binding to the phlO domain, resulting in the expression of GoI, at the time of its concentration exceeds specific value. As Fig.2 shown, while the concentration of 2,4-DAPG is below the threshold, the phlF protein binds to the phlO domain upstream the gene of interest(GOI), which serves as a high order complex and block the recruitment of RNA polymerase, leading to the inhibition of GoI expression. Therefore, we can use DAPG, phlO and phlF to construct a new type of transcriptional switch. ![]() PhlF protein bound its operator sequence (phlO) in the absence of 2,4-diacetylphloroglucinol (DAPG) but dissociated from phlO in the presence of DAPG. Further study shows that phlF gene codes for a repressor protein which can interact with an inverted repeated sequence, phlO, located upstream of the gene related to biosynthesis of DAPG. Recently, it is demonstrated that 2,4-DAPG can serve as signal molecule to construct a transcriptional switch that is similar to Tet on/off system.Īccording to a research conducted to study the biosynthesis pathway of DAPG, researchers found that there is a negative correlation between phlF gene expression level and the DAPG synthetic amount. Introduction-A New Type of Transcriptional SwitchĢ,4-DAPG (DAPG), full name 2,4-diacetylphloroglucinol, is a natural phenol found in specific strains of Gram-negative bacterium Pseudomonas fluorescens. Luckily, after further literature reading and brainstorming, we find this system will bring us more surprise. atroviride will respond to the invasion of pathogens. atroviride through the DAPG-Off/On system, and T. When our device detected that the plant is infected with pathogens and is in poor condition by analyzing the changes in volatile organic compounds (VOC) released by plants, it will tell our engineered T. We choose the DAPG-Off/On System to serve as the link between our device and engineered organisms for it is supersensitive. Hence it requires us to make our device and our engineered organisms connected. This year, we aim to construct a diseases-monitoring system for automatic agriculture in which hardware and organisms can act as a whole. ![]()
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