Expression Analysis Reveals That Sorghum Disease Resistance Protein SbSGT1 Is Regulated by Auxin

SGT1 (suppressor of the skp1 G2 allele) is an important plant disease resistance-related protein, which plays an important role in plant resistance to pathogens and regulates signal transduction during the process of plant disease resistance. In this study, we analyzed the expression profile of SbSGT1 in sorghum under phytohormones treatment. Quantitative real-time PCR results showed that SbSGT1 was most expressed in sorghum leaves, and could respond to plant hormones such as auxin, abscisic acid, salicylic acid, and brassinolide. Subsequently, we determined the optimal soluble prokaryotic expression conditions for SbSGT1 and purified it using a protein purification system in order to evaluate its potential interactions with plant hormones. Microscale thermophoretic analysis showed that SbSGT1 exhibited significant interactions with indole-3-acetic acid (IAA), with a Kd value of 1.5934. Furthermore, the transient expression of SbSGT1 in Nicotiana benthamiana indicated that treatment with exogenous auxin could inhibit SbSGT1 expression, both at the transcriptional and translational level, demonstrating that there exists an interaction between SbSGT1 and auxin.

Gold Nanoparticles Augment N-Terminal Cleavage and Splicing Reactions in Mycobacterium tuberculosis SufB

Protein splicing is a self-catalyzed event where the intervening sequence intein cleaves off, joining the flanking exteins together to generate a functional protein. Attempts have been made to regulate the splicing rate through variations in temperature, pH, and metals. Although metal-regulated protein splicing has been more captivating to researchers, metals were shown to only inhibit splicing reactions that confine their application. This is the first study to show the effect of nanoparticles (NPs) on protein splicing. We found that gold nanoparticles (AuNPs) of various sizes can increase the splicing efficiency by more than 50% and the N-terminal cleavage efficiency by more than 45% in Mycobacterium tuberculosis SufB precursor protein. This study provides an effective strategy for engineering splicing-enhanced intein platforms. UV-vis absorption spectroscopy, isothermal titration calorimetry (ITC), and transmission electron microscopy (TEM) confirmed AuNP interaction with the native protein. Quantum mechanics/molecular mechanics (QM/MM) analysis suggested a significant reduction in the energy barrier at the N-terminal cleavage site in the presence of gold atom, strengthening our experimental evidence on heightened the N-terminal cleavage reaction. The encouraging observation of enhanced N-terminal cleavage and splicing reaction can have potential implementations from developing a rapid drug delivery system to designing a contemporary protein purification system.

REVOLVER: a low-cost automated protein purifier based on parallel preparative gravity column workflows

Protein purification is a ubiquitous operation in biochemistry and life sciences and represents a key step to producing purified proteins for research (understanding how proteins work) and various applications. The need for scalable and parallel protein purification systems keeps growing due to the increase in throughput in the production of recombinant proteins and in the ever-growing scale of biochemistry research. Therefore, automating the process to handle multiple samples in parallel with minimal human intervention is highly desirable; yet only a handful of such tools have been developed, all of which are closed source and expensive. To address this challenge, we present REVOLVER, a 3D-printed programmable and automatic protein purification system based on gravity-column workflows and controlled by Arduino boards that can be built for under $130 USD. REVOLVER completes a full protein purification process with almost no human intervention and yields results equivalent to those obtained by an experienced biochemist when purifying a real-world protein sample. We further present and describe MULTI-VOLVER, a scalable version of the REVOLVER that allows for parallel purification of up to six samples and can be built for under $250 USD. Both systems will be useful to accelerate protein purification and ultimately link them to bio-foundries for protein characterization and engineering.

Case Study of Selected Nature Swimming Pools in the South Moravian Region

Nature swimming pools are a sought-after alternative to conventional swimming pools. Their natural water purification system, without the need for chemicals, is beneficial to health and environmentally friendly. Designing such a nature swimming pool is a complex matter because it requires coordination of a number of effects, which can easily cause an imbalance in the entire system. This work uses the case study method to analyse the overall condition of three nature swimming pools in the South Moravian Region during the second half of the swimming pool operation season.

A Field Study for Sustainable Community Empowerment through Appropriate Technology of Water Purification and the Concept of Feces Standard Money in Hatphain Village, Lao PDR

The application of appropriate technologies in a developing country is an opportunity to introduce green technology which may lead people to imagine a better life. This point is at the intersection of appropriate technology and sustainable development. This research focuses on Hatphain village where there is no clean water, sewage system, or power lines. Two environmental technologies (an unpowered water purification system, Ongdalsaem and an eco-toilet system, BeeVi toilet) were introduced, based on the concepts of the environmental self- sustainable village and feces Standard Money (fSM). We found that the Ongdalsaem was effective in reducing the concentration of nitrogen and lead in the water in Hatphain. The BeeVi water-free toilet was installed, allowing urine and feces to be separated and used as compost. We strived to spread ideas of sustainability using new eco-circulation experiences and encouraging learning about environmental technology through practical and playing facilities that residents managed themselves.