Mycoplasma hyopneumoniae, the etiological agent of swine enzootic pneumonia, is an important pathogen in the swine industry worldwide. Vaccination is the most cost-effective strategy for controlling and prevention of this disease. However, investigations on pathogenicity mechanisms as well as current serological detection methods and the development of new recombinant subunit vaccines are hampered by the lack of known and well characterized species-specific M. hyopneumoniae antigens. In this work, 54 predicted genes encoding proteins with potential to be used as subunit vaccine or antigens in diagnostic tests were selected, amplified by PCR and cloned into Escherichia coli expression vectors. Recombinant protein expression, solubility and yields were analyzed. The majority of the recombinant proteins were expressed in inclusion bodies. After solubilization with urea or N-lauroyl sarcosine, recombinant proteins were purified by Ni(2+) affinity chromatography. This approach allowed purification of thirty recombinant M. hyopneumoniae proteins which will be evaluated as vaccine candidates and/or as antigens to be used in diagnostic tests.
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http://dx.doi.org/10.1016/j.pep.2009.09.001 | DOI Listing |
Int J Biol Macromol
January 2025
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China; Zhaoqing Branch Centre of Guangdong Laboratory for Lingnan Modern Agricultural Science and Technology, Zhaoqing 526238, China; Zhaoqing Institute of Biotechnology Co., Ltd., Zhaoqing 526238, China; Guangdong Wens Dahuanong Bio-Pharmaceutical Co., Ltd., Xinxing 527400, China. Electronic address:
Virus-host protein interaction is critical for successful completion of viral replication cycles. As the largest nonstructural protein (NSP) of porcine reproductive and respiratory syndrome virus (PRRSV), NSP2 plays multiple and critical roles in viral replication, antiviral immunity, cellular tropism and virulence. An interactome of this protein with host proteins would be instrumental in full understanding of these multifunctional roles.
View Article and Find Full Text PDFJ Biotechnol
January 2025
Institute of Molecular Biotechnology (IMBT), BOKU University, Vienna, Austria.
Efficient recombinant protein production requires mammalian stable cell lines or often relies on inefficient transfection processes. Baculoviral transduction of mammalian cells (BacMam) offers cost-effective and robust gene transfer and straightforward scalability. The advantages over conventional approaches are, no need of high biosafety level laboratories, efficient transduction of various cell types and transfer of large transgenes into host cells.
View Article and Find Full Text PDFSTAR Protoc
January 2025
Department of Chemistry, School of Science, Westlake University, Hangzhou, Zhejiang Province 310030, China; Institute of Natural Sciences, Westlake Institute for Advanced Study, Hangzhou, Zhejiang Province 310024, China; Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang Province 310024, China. Electronic address:
Post-translational modifications (PTMs) of histone H4 play significant roles in the regulation of chromatin status. Here, we present a protocol for semisynthesis of histone H4 by sortase-mediated ligation (SML). We describe steps for solid-phase peptide synthesis of H4R40C(1-42), recombinant expression and purification of H4(41-102), expression and purification of eSrt(2A-9), and preparation of acrylamidine.
View Article and Find Full Text PDFFEBS J
January 2025
Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Gothenburg, Sweden.
Mucus in the colon is crucial for intestinal homeostasis by forming a barrier that separates microbes from the epithelium. This is achieved by the structural arrangement of the major mucus proteins, such as MUC2 and FCGBP, both of which are comprised of several von Willebrand D domains (vWD) and assemblies. Numerous disulfide bonds stabilise these domains, and intermolecular bonds generate multimers of MUC2.
View Article and Find Full Text PDFProbiotics Antimicrob Proteins
January 2025
Faculty of Biotechnologies (BioTech), ITMO University, 9 Lomonosova Street, 191002, Saint Petersburg, Russia.
Antimicrobial peptides (AMPs) are small, positively charged biomolecules produced by various organisms such as animals, microbes, and plants. These AMPs play a significant role in defense mechanisms and protect from adverse conditions. The emerging problem of drug resistance in microbes poses a global health challenge in treating diseases.
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