Green fluorescent protein (GFP) is commonly used as a reporter protein in a wide range of biological experiments. The efficient protocol of Agrobacterium-mediated transient expression in Nicotiana excelsior was applied for quick preparative production of recombinant GFP. The protein purification scheme has been developed and included ammonium sulfate precipitation and Q-sepharose anion-exchange chromatography. It results in obtaining of a fraction with about 85% GFP homogeneity and the protein yield of about 75%.
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BioTech (Basel)
January 2025
Federal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, pr. Lavrentieva 10, Novosibirsk 630090, Russia.
The development of efficient producers of recombinant pharmaceuticals based on plant cell suspension cultures is a pressing challenge in modern applied science. A primary limitation of plant cell cultures is their relatively low yield of the target protein. One strategy to enhance culture productivity involves reducing cell aggregation.
View Article and Find Full Text PDFGene Ther
January 2025
Nantes Université, CHU Nantes, INSERM, TaRGeT-Translational Research in Gene Therapy, UMR1089, F-44200, Nantes, France.
The liver is a unique organ where immunity can be biased toward ineffective response notably in the context of viral infections. Chronic viral hepatitis depends on the inability of the T-cell immune response to eradicate antigen. In the case of recombinant Adeno-Associated-Virus, used for therapeutic gene transfer, conflicting reports describe tolerance induction to different transgene products while other studies have shown conventional cytotoxic CD8 T cell responses with a rapid loss of transgene expression.
View Article and Find Full Text PDFJ Org Chem
January 2025
Department of Chemistry, The University of Texas at Austin, Austin, Texas 78712, United States.
Green fluorescent protein (GFP) chromophores are widely studied as fluorescent moieties for sensing and imaging applications. Herein, we present a straightforward synthetic strategy that involves the reaction of glycine amides with 1,3-diketones to form imidazolones through an unusual molecular fragmentation and recombination pathway. Mechanistic investigations, including crossover experiments, inspired a competing strategy that incorporates exogenous ketones into the products, yielding fluorescent GFP chromophore analogues.
View Article and Find Full Text PDFFront Immunol
January 2025
Institute of Biopharmaceutical and Health Engineering, Tsinghua Shenzhen International Graduate School, Tsinghua University, Shenzhen, China.
Background: Epstein-Barr virus (EBV) is a significant global public health concern because of its association with various malignancies and autoimmune diseases. Over 90% of the global population is chronically infected with EBV, impacting numerous cancer-related cases annually. However, none of the effective prophylactic vaccines against EBV is approved at present.
View Article and Find Full Text PDFAppl Microbiol Biotechnol
January 2025
Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Av. Instituto Politécnico Nacional No, 2508, C.P. 07360, Mexico City, Mexico.
One of the most significant bacteriophage technologies is phage display, in which heterologous peptides are exhibited on the virion surface. This work describes the display of λ decorative protein D linked to the E protein domain III of Zika virus (D-ZE), to the GFP protein (D-GFP), or to different domain III epitopes of the E protein (D-TD), exhibited on the surface of an in vitro evolved lambda phage (λ). This phage harbors a gene D deletion and was subjected to directed evolution using Escherichia coli W3110/pD-ZE as background.
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