Self-cleaving peptides for expression of multiple genes in Dictyostelium discoideum.

The social amoeba Dictyostelium discoideum is a model for a wide range of biological processes including chemotaxis, cell-cell communication, phagocytosis, and development. Interrogating these processes with modern genetic tools often requires the expression of multiple transgenes. While it is possible to transfect multiple transcriptional units, the use of separate promoters and terminators for each gene leads to large plasmid sizes and possible interference between units. In many eukaryotic systems this challenge has been addressed through polycistronic expression mediated by 2A viral peptides, permitting efficient, co-regulated gene expression. Here, we screen the most commonly used 2A peptides, porcine teschovirus-1 2A (P2A), Thosea asigna virus 2A (T2A), equine rhinitis A virus 2A (E2A), and foot-and-mouth disease virus 2A (F2A), for activity in D. discoideum and find that all the screened 2A sequences are effective. However, combining the coding sequences of two proteins into a single transcript leads to notable strain-dependent decreases in expression level, suggesting additional factors regulate gene expression in D. discoideum that merit further investigation. Our results show that P2A is the optimal sequence for polycistronic expression in D. discoideum, opening up new possibilities for genetic engineering in this model system.

Potential use of only Yb2O3 in producing dense Si3N4 ceramics with high thermal conductivity by gas pressure sintering.

Yb2O3 is an efficient sintering additive for enhancing not only thermal conductivity but also the high-temperature mechanical properties of Si3N4 ceramics. Here we report the fabrication of dense Si3N4 ceramics with high thermal conductivity by the gas pressure sintering of α-Si3N4 powder compacts, using only Yb2O3 as an additive, at 1900 °C under a nitrogen pressure of 1 MPa. The effects of Yb2O3 content, sample packing condition and sintering time on the densification, microstructure and thermal conductivity were investigated. Curves of the density plotted against the Yb2O3 content exhibited a characteristic 'N' shape with a local minimum at 3 mol% Yb2O3 and nearly complete densification below and above this concentration. The effects of the sample packing condition on the densification, microstructure and thermal conductivity strongly depended on the Yb2O3 content. The embedded condition led to more complete densification but also to a decrease in thermal conductivity from 119 to 94 W m-1 K-1 upon 1 mol% Yb2O3 addition. The sample packing condition had little effect on the density and thermal conductivity (102-106 W m-1 K-1) at 7 mol% Yb2O3. The thermal conductivity value was strongly related to the microstructure.