We describe the expression of recombinant fluorescent proteins in the milk of two lines of transgenic pigs generated by Sleeping Beauty transposon-mediated genetic engineering. The Sleeping Beauty transposon consisted of an ubiquitously active CAGGS promoter driving a fluorophore cDNA, encoding either Venus or mCherry. Importantly, the fluorophore cDNAs did not encode for a signal peptide for the secretory pathway, and in previous studies of the transgenic animals a cytoplasmic localization of the fluorophore proteins was found. Unexpectedly, milk samples from lactating sows contained high levels of bioactive Venus or mCherry fluorophores. A detailed analysis suggested that exfoliated cells of the mammary epithelium carried the recombinant proteins passively into the milk. This is the first description of reporter fluorophore expression in the milk of livestock, and the findings may contribute to the development of an alternative concept for the production of bioactive recombinant proteins in the udder.
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http://dx.doi.org/10.1038/srep24464 | DOI Listing |
Molecular rotor-based fluorophores (RBFs) that are target-selective and sensitive to both polarity and viscosity are valuable for diverse biological applications. Here, we have designed next-generation RBFs based on the underexplored bimane fluorophore through either changing in aryl substitution or varying π-linkages between the rotatable electron donors and acceptors to produce red-shifted fluorescence emissions with large Stokes shifts. RBFs exhibit a twisted intramolecular charge transfer mechanism that enables control of polarity and viscosity sensitivity, as well as target selectivity.
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January 2025
Translational Glycomics Research Group, Research Institute of Biomolecular and Chemical Engineering, University of Pannonia, Veszprem, Hungary; Horváth Csaba Memorial Laboratory of Bioseparation Sciences, Research Center for Molecular Medicine, Faculty of Medicine, University of Debrecen, Hungary. Electronic address:
Sodium dodecyl sulfate capillary gel electrophoresis (SDS-CGE) is a frequently used analytical technique in size-based separation of proteins, playing a vital role in the biopharmaceutical industry for the analysis and characterization of therapeutic proteins, employing both UV and fluorescent detection. Understanding the effect of the operational parameters using easily applicable in migratio fluorescent labeling is increasingly critical, especially because multicapillary electrophoresis systems with fluorescent detection have recently gained prominence in high-throughput biopolymer analysis. In this study, the effects of the three most important user-adjustable operational parameters (temperature, gel concentration, and electric field strength) were investigated on the electrophoretic mobility and resolution of SDS-protein complexes in the presence of propidium iodide in the gel-buffer system.
View Article and Find Full Text PDFNat Commun
January 2025
Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Germany.
HIV-associated neurocognitive disorders (HAND) and viral reservoirs in the brain remain a significant challenge. Despite their importance, the mechanisms allowing HIV-1 entry and replication in the central nervous system (CNS) are poorly understood. Here, we show that α-synuclein and (to a lesser extent) Aβ fibrils associated with neurological diseases enhance HIV-1 entry and replication in human T cells, macrophages, and microglia.
View Article and Find Full Text PDFRSC Chem Biol
January 2025
Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences Flemingovo n. 2 Prague 6 Czechia
The LutR protein represses the transcription of genes encoding enzymes for the utilization of l-lactate in through binding to a specific DNA region. In this study, we employed oligonucleotide probes modified by viscosity-sensitive tetramethylated thiophene-BODIPY fluorophores to investigate the impact of selected metabolites on the LutR-DNA complex. Our goal was to identify the effector molecule whose binding alters the protein-DNA affinity, thereby enabling gene transcription.
View Article and Find Full Text PDFSci Rep
January 2025
Department of Veterinary Anatomy, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo, 113-8657, Japan.
An aqueous solution of a common food dye, Fast Green FCF (FG), mimics cholyl-lysyl-fluorescein to visualize embryonic bile flow via single peritoneal injection into intrauterine mouse embryos. Despite its efficacy in embryos, its suitability for adult mice and small to medium-sized mammals remained uncertain. In this study, we investigated FG cholangiography in adult mice, dogs, and goats.
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