Browsing by Author "Frank, K."

Now showing 1 - 2 of 2
  • Thumbnail Image
    Item
    In vitro and in vivo detection of microbial gene expression in bioactivated scaffolds seeded with cyanobacteria
    (WILEY, 2022) Leibrock, L. B.; Hofmann, D. M.; Fuchs, B.; Birt, A.; Reinholz, M.; Guertler, A.; Frank, K.; Giunta, R. E.; Egaña Erazo Jose Tomas; Nickelsen, J.; Schenck, T. L.; Moellhoff, N.
    Dermal replacement materials bioactivated with cyanobacteria have shown promising potential for wound regeneration. To date, extraction of cyanobacteria RNA from seeded scaffolds has not been described. The aim of this study was to develop a method to isolate total RNA from bioactivated scaffolds and to propose a new approach in determining living bacteria based on real-time PCR. Transgenic Synechococcus sp. PCC 7002 (tSyn7002) were seeded in liquid cultures or scaffolds for dermal regeneration in vitro and in vivo for 7 days. RNA was extracted with a 260/280 ratio of >= 2. The small subunit of the 30S ribosome in prokaryotes (16S) and RNAse P protein (rnpA) were validated as reference transcripts for PCR analysis. Gene expression patterns differed in vitro and in vivo. Expression of 16S was significantly upregulated in scaffolds in vitro, as compared to liquid cultures, whilst rnpA expression was comparable. In vivo, both 16S and rnpA showed reduced expression compared to in vitro (16S: in vivo Ct value 13.21 +/- 0.32, in vitro 12.44 +/- 0.42; rnpA in vivo Ct value 19.87 +/- 0.41, in vitro 17.75 +/- 1.41). Overall, the results demonstrate rnpA and 16S expression after 7 days of implantation in vitro and in vivo, proving the presence of living bacteria embedded in scaffolds using qPCR.
  • No Thumbnail Available
    Item
    Structure-based characterization of multiprotein complexes
    (2014) Wiederstein, M.; Gruber, M.; Frank, K.; Melo Ledermann, Francisco Javier; Sippl, M. J.
    Multiprotein complexes govern virtually all cellular processes. Their 3D structures provide important clues to their biological roles, especially through structural correlations among protein molecules and complexes. The detection of such correlations generally requires comprehensive searches in databases of known protein structures by means of appropriate structure-matching techniques. Here, we present a high-speed structure search engine capable of instantly matching large protein oligomers against the complete and up-to-date database of biologically functional assemblies of protein molecules. We use this tool to reveal unseen structural correlations on the level of protein quaternary structure and demonstrate its general usefulness for efficiently exploring complex structural relationships among known protein assemblies.