A method is described for using 96-well plates to prepare libraries of Escherichia coli cultures for screening a library of gene variants. This approach bypasses colony-picking to allow standard molecular biology laboratories to carry out directed evolution efficiently with a 96-well plate-reader and multichannel pipettes. Initial screens are applied to cultures that are rapidly prepared by diluting transformed cells so that an average of four cells starts each culture. Subsequent screens are used to isolate individual enzyme-expressing clones that exhibit activity higher than the parental clone. The outlined method also includes guidelines for preparing a library of gene variants and for optimizing a screening method.
College of Materials, Institute of Artificial Intelligence, State Key Laboratory of Physical Chemistry of Solid Surfaces, College of Chemistry and Chemical Engineering, iChEM, Fujian Key Laboratory of Advanced Materials, College of Energy, Xiamen University, Xiamen 361005, China.
Nickel-iron-based catalysts are recognized for their high efficiency in the oxygen evolution reaction (OER) under alkaline conditions, yet the underlying mechanisms that drive their superior performance remain unclear. Herein, we revealed the molecular OER mechanism and the structure-intermediate-performance relationship of OER on a phosphorus-doped nickel-iron nanocatalyst (NiFeP). NiFeP exhibited exceptional activity and stability with an overpotential of only 210 mV at 10 mA cm in 1 M KOH and a cell voltage of 1.
Department of Biological and Environmental Sciences, Faculty of Natural Sciences, Walter Sisulu University, Nelson Mandela Drive Campus, P/Bag X1, Mthatha 5117, Eastern Cape, South Africa.
Invertebrates form a vital component of agricultural ecosystems, and they are chief actors in sustaining the functions of the ecosystem and soil health. Scholarly publications that concentrated on visualizing the research outputs and trends on invertebrates and agriculture are scarce. In this paper, we adopted a bibliometric model to extract trends/research studies on invertebrates and agriculture between 1991 and 2022, using scholarly studies retrieved from the Web of Science (WoS) databank.
Sweetpotato (Ipomoea batatas L.), an important food and industrial crop in the world, has a highly heterozygous hexaploid genome, making the development of single nucleotide polymorphism (SNP) markers challenging. Identifying SNP loci and developing practical SNP markers are crucial for genomic and genetic research on sweetpotato.
Sequence-function data provides valuable information about the protein functional landscape but is rarely obtained during directed evolution campaigns. Here, we present Long-read every variant Sequencing (LevSeq), a pipeline that combines a dual barcoding strategy with nanopore sequencing to rapidly generate sequence-function data for entire protein-coding genes. LevSeq integrates into existing protein engineering workflows and comes with open-source software for data analysis and visualization.
AbstractTheoretical studies from diverse areas of population biology have shown that demographic stochasticity can substantially impact evolutionary dynamics in finite populations, including scenarios where traits that are disfavored by natural selection can nevertheless increase in frequency through the course of evolution. Here, we analytically describe the eco-evolutionary dynamics of finite populations from demographic first principles. We investigate how noise-induced effects can alter the evolutionary fate of populations in which total population size may vary stochastically over time.
