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    Tetrahymena thermophila granule lattice protein 3 improves solubility of sexual stage malaria antigens expressed in Escherichia coli
    (Academic Press Inc Elsevier Science, 2022) Akkale, Cengiz; Cassidy-Hanley, Donna Marie; Clark, Theodore G.
    The requirement for low cost manufacturing makes bacterial cells a logical platform for the production of recombinant subunit vaccines for malaria. However, protein solubility has been a major stumbling block with prokaryotic expression systems. Notable examples include the transmission blocking vaccine candidates, Pfs25 and Pfs48/45, which are almost entirely insoluble when expressed as recombinant proteins in Escherichia coli. Various solubility tags have been used with limited success in improving solubility, although recent studies with granule lattice protein 1 (Grl1p) from the ciliated protozoan, Tetrahymena thermophila, have shown promise. Here, we examine a related solubility tag, granule lattice protein 3 (Grl3p) from T. thermophila, and compare it to both Grl1p and the well-studied maltose binding protein (MBP) used to improve the solubility of multiple protein targets. We find that Grl3p performs comparably to Grl1p when linked to Pfs25 but significantly improves solubility when paired with Pfs48/45.
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    Öğe
    The use of CRISPR-Cas-based systems in bacterial cell factories
    (Elsevier, 2023) Kolasinliler, Gulsen; Aagre, Mehdi Movahed; Akkale, Cengiz; Kaya, Hilal Betul
    Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas system has been the focus since its first introduction to genome editing research. It acts as an adaptive immune system in many prokaryotes by detecting and degrading viral/invasive DNA/RNA repurposed for targeted genome editing in various organisms, including bacterial species relevant to industrial biotechnology. Targeted genome editing in bacteria has been implemented for multiple purposes, such as method development, understanding knowledge of metabolic processes and improving the productivity of industrial products. With the increasing share of biotechnological products, bac-teria have been the subject in the field of genome editing.This review summarizes the immunological mechanisms of CRISPR-Cas systems, the variation in classification of CRISPR-Cas systems by highlighting their features and the current applications of CRISPR-based genome editing in industrially relevant bacteria. In addition, an extensive literature review focused on recent advances has been presented in bacterial species according to industrial significance.