The bead transfection method involves binding nucleic acids onto 3-microm-diameter paramagnetic beads, treating the beads with transfection reagent, and using them as scaffolds to direct transfection to individual cells or regions in a population. Typically, PCR products are used because they can be conveniently generated using biotinylated primers and can introduce site-directed mutations, without the need for cloning or plasmid purification. However, the method can be adapted to transfect plasmid DNA or RNA. The magnetic properties of the beads allows magnets to direct the loci of transfection in cell culture; magnetic arrays are built in cell culture chambers to allow multiple parallel transfections on the same microscope coverslip. The PCR reaction and transfection can be carried out in 1 d, and transfection results can be viewed in 24-48 h.
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http://dx.doi.org/10.1038/nprot.2006.74 | DOI Listing |
Int J Biol Macromol
January 2025
Department of Applied Biology, School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China.
Lincomycin, produced by the actinomycete Streptomyces lincolnensis, is highly effective against Gram-positive bacteria and protozoans, making it widely used in clinical settings. This study identified LcbR2, a MarR family transcriptional regulator, as an activator of lincomycin biosynthesis. Knocking out the lcbR2 gene reduced lincomycin production by 63.
View Article and Find Full Text PDFJ Biol Chem
January 2025
Rosalind and Morris Goodman Cancer Institute, McGill University, Montreal, Quebec H3A 1A3, Canada; Department of Medicine, McGill University, Montreal, Quebec H3A 1A3, Canada; Department of Biochemistry, McGill University, Montreal, Quebec H3A 1A3, Canada; McGill University Health Center, Montreal, Quebec H3A 1A3, Canada. Electronic address:
Site-directed mutagenesis is a fundamental tool indispensable for protein and plasmid engineering. An important technological question is how to achieve the efficiency at the ideal level of 100%. Based on complementary primer pairs, the QuickChange method has been widely used, but it requires significant improvements due to its low efficiency and frequent unwanted mutations.
View Article and Find Full Text PDFGene
January 2025
Jiangxi Provincial Key Laboratory of Poultry Genetic Improvement, Nanchang 330032 China. Electronic address:
The growth and development of chicken skeletal muscle directly affects chicken meat production, which is very important for broiler industry. Matrix metallopeptidase 2 (MMP2) exists in skeletal muscle. However, the underlying regulating of MMP2 remain unknown.
View Article and Find Full Text PDFMicrob Pathog
January 2025
Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Ministry of Education, International Research Center for Marine Biosciences at Shanghai Ocean University, Ministry of Science and Technology, National Demonstration Center for Experimental Fisheries Science Education, Shanghai Ocean University, Shanghai 201306, China; Marine Biomedical Science and Technology Innovation Platform of Lin-gang Special Area, Shanghai 201306, China. Electronic address:
Vibrio anguillarum is a pathogen responsible for vibriosis in aquaculture animals. The formation of bacterial biofilm contributes to infections and increases resistance to antibiotics. Tryptophanase and its substrate tryptophan have been recognized as signal molecules regulating bacterial biofilm formation.
View Article and Find Full Text PDFFood Chem Toxicol
January 2025
Department of Molecular and Translational Medicine, University of Brescia, Italy.
Background: Methylglyoxal (MGO), a highly reactive precursor of advanced glycation end products (AGEs), is endogenously produced and prevalent in various ultra-processed foods. MGO has emerged as a significant precursor implicated in the pathogenesis of type 2 diabetes and neurodegenerative diseases. To date, the effects of dietary MGO on the intestine have been limited explored.
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