Frank Wilco Bartels

Specific binding of the regulatory protein ExpG to promoter regions of the galactoglucan biosynthesis gene cluster of Sinorhizobium meliloti - a combined molecular biology and force spectroscopy investigation

Specific protein-DNA interaction is fundamental for all aspects of gene transcription. We focus on a regulatory DNA-binding protein in the Gram-negative soil bacterium Sinorhizobium meliloti 2011, which is capable of fixing molecular nitrogen in a symbiotic interaction with alfalfa plants. The ExpG protein plays a central role in regulation of the biosynthesis of the exopolysaccharide galactoglucan, which promotes the establishment of symbiosis. ExpG is a transcriptional activator of exp gene expression. We investigated the molecular mechanism of binding of ExpG to three associated target sequences in the exp gene cluster with standard biochemical methods and single molecule force spectroscopy based on the atomic force microscope (AFM). Binding of ExpG to expA1, expG-expD1, and expE1 promoter fragments in a sequence specific manner was demonstrated, and a 28 bp conserved region was found. AFM force spectroscopy experiments confirmed the specific binding of ExpG to the promoter regions, with unbinding forces ranging from 50 to 165 pN in a logarithmic dependence from the loading rates of 70-79000 pN/s. Two different regimes of loading rate-dependent behaviour were identified. Thermal off-rates in the range of k(off)=(1.2+/-1.0) x 10(-3)s(-1) were derived from the lower loading rate regime for all promoter regions. In the upper loading rate regime, however, these fragments exhibited distinct differences which are attributed to the molecular binding mechanism.

Covalent and physical cross-linking of photonic crystals with 10-fold-enhanced chemomechanical stability.

We report opal photonic crystals that are self-assembled from functional polymer particles. We randomly copolymerized functional side-chain monomers containing motifs that form homodimers or heterobridges. These include ether or methylene bridges, hydrazone bridges, acids for anhydride formation, low- T g copolymers or physical cross-links by hydrogen bonds and/or polarity. To generate particles that are monodisperse, spherical, and functionalized, we combined emulsifier-free synthesis with swelling synthesis steps. Laser diffraction from centimeter-sized beams, white-light interferometry, and atomic force microscopy demonstrates symmetry and homogeneity across the entire crystal without the loss of interstitial volume. Compared to the stability of nonfunctional particles, the stability of the crystal against immersion in water and isopropanol was enhanced from 10 to a perfect 100%. One of the successful approaches (methylene bridges from N-methylolmethacrylamid) is triggered by thermal activation, but as shown, this is operative far from the trivial regime of sintering. We demonstrate successful infiltration with and solvation of a laser-polymerizable resin, thus enabling the processing of 3D photonic waveguide structures.

Tunable, elastic, crack-free photonic crystals and polymer opal templates with pre-determined orientation for defect inscription

Self-assembled polymer opal colloidal crystals can be useful either as templates for processing, followed by inversion with high-index material (leading to waveguide structures inside a full-band-gap photonic crystal) or as a stop-band material that has special mechanical properties, not achievable with the inorganic high-index-materials. Both options are explored in the present contribution.