While clathrin heavy chains from different species are highly conserved in amino acid sequence, clathrin light chains are much more divergent. Thus clathrin light chain may have different functions in different organisms. To investigate clathrin light chain function, we cloned the clathrin light chain, clcA, from Dictyostelium and examined clathrin function in clcA-mutants. Phenotypic deficiencies in development, cytokinesis, and osmoregulation showed that light chain was critical for clathrin function in Dictyostelium. In contrast with budding yeast, we found the light chain did not influence steady-state levels of clathrin, triskelion formation, or contribute to clathrin over-assembly on intracellular membranes. Imaging GFP-CHC in clcA-mutants showed that the heavy chain formed dynamic punctate structures that were remarkably similar to those found in wild-type cells. However, clathrin light chain knockouts showed a decreased association of clathrin with intracellular membranes. Unlike wild-type cells, half of the clathrin in clcAmutants was cytosolic, suggesting that the absence of light chain compromised the assembly of triskelions onto intracellular membranes. Taken together, these results suggest a role for the Dictyostelium clathrin light chain in regulating the self-assembly of triskelions onto intracellular membranes, and demonstrate a crucial contribution of the light chain to clathrin function in vivo.

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Epitaxial p-i-n structures grown on native GaN substrates have been fabricated and used to extract the impact ionization coefficients in GaN. The photomultiplication method has been used to experimentally determine the impact ionization coefficients; avalanche dominated breakdown is confirmed by variable-temperature breakdown measurements. To facilitate photomultiplication measurements of both electrons and holes, the structures include a thin pseudomorphic In0.07Ga0.93N layer on the cathode side of the drift layer. Illumination with 193 nm and 390 nm UV light has been performed on diodes with different intrinsic layer thicknesses. From the measured multiplication characteristics, the impact ionization coefficients of electrons (α) and holes (β) were determined for GaN over the electric field range from 2 MV/cm to 3.7 MV/cm. The results show that for transport along the c-axis, holes dominate the impact ionization process at lower electric field strengths; the impact ionization coefficient of electrons becomes comparable to that of holes (β/α<5) for electric field strengths above 3.3 MV/cm.

I- CreI is a member of the LAGLI-DADG family of homing nucleases; however, unlike most members of this family it contains only a single copy of this signature motif. I- CreI was over-expressed in Escherichia coli, and a simple purification protocol developed that gave reasonably pure protein in high yield. Size-exclusion chromatography and chemical cross-linking indicated that the protein is a dimer in solution. DNA cleavage by I- CreI was absolutely dependent on Mg2+(or Mn2+), and was inhibited by monovalent cations. I- CreI displayed a surprisingly high temperature optimum (>50 degrees C), with full activity occurring even at 70 degrees C. Interestingly, SDS was needed for efficient release of the cleavage products from the protein, indicating formation of very stable DNA-protein complexes. In contrast to these robust characteristics, purified I- CreI was unstable; however, it could be stabilized by the addition of either target or non-target DNA. Mobility shift assays revealed that I- CreI binds to DNA in the absence of Mg2+. Hydroxyl radical footprinting showed that I- CreI strongly protected the backbone of a continuous stretch of at least 12 nt on each strand that were shifted, relative to each other, by 2 bp in the 3'direction. Methylation protection and interference analyses were also performed, and together with the hydroxyl radical footprinting, indicate that I- CreI binds in both the major and minor grooves of its target DNA.

An epitaxial lift‐off (ELO) process for GaN materials has been demonstrated using bandgap‐selective photoenhanced wet etching of an InGaN release layer. This process has been applied to GaN layers grown on sapphire as well as native GaN substrates using a perforation technique to scale the process up to wafers of arbitrary size. The process has the advantage of leveraging conventional MOCVD growth to form the release layer, with minimal degradation of films grown on top of the release layer. The ELO process is non‐destructive and can enable cost reduction through reuse of the native GaN substrate after ELO. The GaN films have been characterized before and after ELO using AFM, SEM, XRD, TEM and by fabricating Schottky barrier diodes. The performance of Schottky diodes fabricated on GaN‐on‐sapphire structures was found to improve after ELO. Potential applications for this technology include GaN power and optoelectronic devices as well as flexible electronics. Shown is a 5‐micron‐thick GaN epitaxial film released from a 4‐inch sapphire substrate using perforations on a 1‐mm pitch. The yellow luminescence of the nitrogen face of the released film is visible under ultraviolet illumination.

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