Most damage induced mutagenesis in Escherichia coli is dependent upon the UmuD'(2)C protein complex, which comprises DNA polymerase V (pol V). Biochemical characterization of pol V has been hindered by the fact that the enzyme is notoriously difficult to purify, largely because overproduced UmuC is insoluble. Here, we report a simple and efficient protocol for the rapid purification of milligram quantities of pol V from just 4 L of bacterial culture. Rather than over producing the UmuC protein, it was expressed at low basal levels, while UmuD'(2)C was expressed in trans from a high copy-number plasmid with an inducible promoter. We have also developed strategies to purify the β-clamp and γ-clamp loader free from contaminating polymerases. Using these highly purified proteins, we determined the cofactor requirements for optimal activity of pol V in vitro and found that pol V shows robust activity on an SSB-coated circular DNA template in the presence of the β/γ-complex and a RecA nucleoprotein filament (RecA*) formed in trans. This strong activity was attributed to the unexpectedly high processivity of pol V Mut (UmuD'(2)C · RecA · ATP), which was efficiently recruited to a primer terminus by SSB.
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http://dx.doi.org/10.1016/j.dnarep.2012.01.012 | DOI Listing |
Nanomaterials (Basel)
December 2024
School of Materials Science and Engineering, Changchun University of Science and Technology, Changchun 130022, China.
Photocatalytic technology holds significant promise for sustainable development and environmental protection due to its ability to utilize renewable energy sources and degrade pollutants efficiently. In this study, BiOI nanosheets (NSs) were synthesized using a simple water bath method with varying amounts of mannitol and reaction temperatures to investigate their structural, morphological, photoelectronic, and photocatalytic properties. Notably, the introduction of mannitol played a critical role in inducing a transition in BiOI from an n-type to a p-type semiconductor, as evidenced by Mott-Schottky (M-S) and band structure analyses.
View Article and Find Full Text PDFNanomaterials (Basel)
December 2024
Institute of Agricultural Resources and Environment, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China.
With the development of industry, agriculture, and aquaculture, excessive ammonia nitrogen mainly involving ionic ammonia (NH) and molecular ammonia (NH) has inevitable access to the aquatic environment, posing a severe threat to water safety. Photocatalytic technology shows great advantages for ammonia nitrogen removal, such as its efficiency, reusability, low cost, and environmental friendliness. In this study, CP (g-CN/CoP) composite materials, which exhibited high-efficiency ammonia nitrogen removal, were synthesized through a simple self-assembly method.
View Article and Find Full Text PDFJ Fungi (Basel)
November 2024
College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China.
Manganese is one of the trace elements necessary for organisms to maintain normal biological activities and is also a cofactor for manganese superoxide dismutase (Mn-SOD) and manganese peroxidase (MnP). In order to find a simple and effective method to rejuvenate the degenerated strains, we explored the effect of the exogenous addition of MnSO on the antioxidant vigour and productivity of degenerated strains of . The results showed that the exogenous MnSO had no significant effect on the non-degenerated strain T0, but it could effectively increase the mycelial growth rate, mycelial biomass, and LBL decolouring ability of the degenerated strains T10 and T19, and reduce the production cycle and increased the biological efficiency of T10; it helped the severely degenerated T19 to regrow its fruiting body; and it also significantly increased the viability of the matrix-degrading enzymes such as EG, Lac, Xyl, etc.
View Article and Find Full Text PDFJ Funct Biomater
November 2024
Faculty of Biology, Medicine and Health, University of Manchester, Manchester M13 9PL, UK.
Tissue engineering research for neurological applications has demonstrated that biomaterial-based structural bridges present a promising approach for promoting regeneration. This is particularly relevant for penetrating traumatic brain injuries, where the clinical prognosis is typically poor, with no available regeneration-enhancing therapies. Specifically, repurposing clinically approved biomaterials offers many advantages (reduced approval time and achieving commercial scaleup for clinical applications), highlighting the need for detailed screening of potential neuromaterials.
View Article and Find Full Text PDFCurr Issues Mol Biol
December 2024
Molecular Cell Biology, Joseph Gottlieb Kölreuter Institute for Plant Sciences, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany.
Landraces are a critical genetic resource for resilience breeding, offering solutions to prepare agriculture for the challenges posed by climate change. Their efficient utilisation depends on understanding their history and genetic relationships. The current study investigates the phylogenetic relationships of barley landraces from Algeria, varieties from the Near and Middle East, traditional landraces, and modern cultivars from Europe.
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