A5064424937- RNA and protein synthesis mechanisms
- Glycosylation and Glycoproteins Research
- Biochemical and Molecular Research
- Enzyme Structure and Function
- Infant Nutrition and Health
- RNA modifications and cancer
- Protein Structure and Dynamics
- Cancer Research and Treatments
- CRISPR and Genetic Engineering
- Monoclonal and Polyclonal Antibodies Research
- Chemical Synthesis and Analysis
University of Freiburg
2019-2024
Biocrates Life Sciences (Austria)
2024
Abstract To build or dissect complex pathways in bacteria and mammalian cells, it is often necessary to recur at least two plasmids, for instance harboring orthogonal inducible promoters. Here we present SiMPl, a method based on rationally designed split enzymes intein-mediated protein trans -splicing, allowing the selection of cells carrying plasmids with single antibiotic. We show that, compared traditional antibiotics, SiMPl increases production antimicrobial non-ribosomal peptide...
Inteins are proteins that excise themselves out of host and ligate the flanking polypeptides in an auto-catalytic process called protein splicing. In nature, inteins either contiguous or split. case split inteins, two fragments must first form a complex for splicing to occur. Contiguous have previously been artificially because allow distinct applications than ones. Even naturally at unnatural sites obtain with reduced affinity one another, which useful create conditional study...
We recently developed the SiMPl plasmid toolbox, which is constituted by pairs of plasmids, generically indicated as pSiMPlx_N and pSiMPlx_C, can be stably maintained in Escherichia coli with a single antibiotic x. The method exploits split intein gp41–1 to reconstitute enzyme conferring resistance toward x, whereby each fragment expressed from one plasmids pair. pSiMPl are currently available for use ampicillin, kanamycin, chloramphenicol, hygromycin, puromycin. Here, we introduce another...
ABSTRACT We recently developed the SiMPl plasmid toolbox, which is constituted by pairs of plasmids, generically indicated as pSiMPl x _N and _C, can be stably maintained in Escherichia coli with a single antibiotic x. The method exploits split intein gp41-1 to reconstitute enzyme conferring resistance towards x, whereby each fragment expressed from one plasmids pair. are currently available for use ampicillin, kanamycin, chloramphenicol, hygromycin puromycin. Here we introduce another pair...
Abstract Inteins are special proteins that auto-catalytically carry out a protein splicing reaction. Due to their ability post-translationally modify target in vitro and vivo , they used different applications, ranging from purification the construction of Boolean logic gates. So far inteins have been found be either encoded by single gene (contiguous inteins) or two separate ones (split inteins). Previously, it has shown contiguous Ssp Rma DnaB split Npu DnaE intein could artificially three...
Abstract Inteins are proteins that excise themselves out of host and ligate the flanking polypeptides in an auto-catalytic process called protein splicing. They gaining momentum synthetic biology for their ability to post-translationally modify interest. In nature, inteins either contiguous or split, which case two intein fragments must first form a complex splicing occur. So far, heuristic methods have been employed whenever new split site had be identified. To make identification faster,...