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Construction and Characterization of MoClo-Compatible Vectors for Modular Protein Expression in .
ACS Synth Biol
January 2025
Department of Engineering Science, University of Oxford, Oxford OX1 3PJ, U.K.
Cloning methods are fundamental to synthetic biology research. The capability to generate custom DNA constructs exhibiting predictable protein expression levels is crucial to the engineering of biology. Golden Gate cloning, a modular cloning (MoClo) technique, enables rapid and reliable one-pot assembly of genetic parts.
View Article and Find Full Text PDFIn vivo cloning of PCR product via site-specific recombination in Escherichia coli.
Appl Microbiol Biotechnol
June 2024
Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
Over the past years, several methods have been developed for gene cloning. Choosing a cloning strategy depends on various factors, among which simplicity and affordability have always been considered. The aim of this study, on the one hand, is to simplify gene cloning by skipping in vitro assembly reactions and, on the other hand, to reduce costs by eliminating relatively expensive materials.
View Article and Find Full Text PDFA simple and efficient protocol for generating transgenic hairy roots using Agrobacterium rhizogenes.
PLoS One
November 2023
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark.
For decades, Agrobacterium rhizogenes (now Rhizobium rhizogenes), the causative agent of hairy root disease, has been harnessed as an interkingdom DNA delivery tool for generating transgenic hairy roots on a wide variety of plants. One of the strategies involves the construction of transconjugant R. rhizogenes by transferring gene(s) of interest into previously constructed R.
View Article and Find Full Text PDFNovel and efficient method for the reconstruction of adenoviruses through isothermal assembly and its potential applications.
Front Med Technol
January 2023
TechnoVax, Inc., Elmsford, NY, United States.
Adenovirus based vectors are useful tools for vaccine development, gene therapy, and oncolytic virotherapy. Here we describe a novel approach for the genetic engineering of any portion of the adenovirus genome and the reconstruction of either fully replication competent or defective virions. This innovative strategy is rapid, effective and suitable for the manipulation of the entire genome broadening the spectrum of potential applications for the adenovirus system.
View Article and Find Full Text PDFDeletion of the cheZ gene results in the loss of swimming ability and the decrease of adhesion ability to Caco-2 cells in Escherichia coli Nissle 1917.
Folia Microbiol (Praha)
June 2023
School of Life Sciences, Zhaoqing University, Zhaoqing, 526061, China.
The aim of this study was to elucidate the biological functions of the motility regulatory protein CheZ in the probiotic strain Escherichia coli Nissle 1917. A cheZ gene deletion strain Nissle 1917ΔcheZ was constructed using the CRISPR/Cas9 two-plasmid system, and the corresponding complemented strain Nissle 1917ΔcheZ/pBR322-cheZ was established. Combined studies of growth kinetics testing, motility assays, swarming motility assays, and bacterial adherence assays were performed to study the motility regulatory protein CheZ-mediated functions in the prototype Nissle 1917 strain, its isogenic cheZ mutant, and the corresponding complemented strain.
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