Detection and localisation of protein-protein interactions in Saccharomyces cerevisiae using a split-GFP method

An alternative method for monitoring protein-protein interactions in Saccharomyces cerevisiae has been developed. It relies on the ability of two fragments of enhanced green fluorescent protein (EGFP) to reassemble and fluoresce when fused to interacting proteins. Since this fluorescence can be detected in living cells, simultaneous detection and localisation of interacting pairs is possible. DNA sequences encoding N- and C-terminal EGFP fragments flanked by sequences from the genes of interest were transformed into S. cerevisicie JPY5 cells and homologous recombination into the genome verified by PCR. The system was evaluated by testing known interacting proteins: labelling of the phosphofructokinase subunits, Pfk1p and Pfk2p, with N- and C-terminal EGFP fragments, respectively, resulted in green fluorescence in the cytoplasm. The system works in other cellular compartments: labelling of Idh1p and Idh2p, (mitochondrial matrix), Sdh3p and Sdh4p (mitochondrial membrane) and Pap2p and Mtr4p (nucleus) all resulted in fluorescence in the appropriate cellular compartment. (c) 2008 Elsevier Inc. All rights reserved.

Simulation or measurement: the effect of radio map creation on indoor WLAN-based localisation accuracy

Indoor wireless network based client localisation requires the use of a radio map to relate received signal strength to specific locations. However, signal strength measurements are time consuming, expensive and usually require unrestricted access to all parts of the building concerned. An obvious option for circumventing this difficulty is to estimate the radio map using a propagation model. This paper compares the effect of measured and simulated radio maps on the accuracy of two different methods of wireless network based localisation. The results presented indicate that, although the propagation model used underestimated the signal strength by up to 15 dB at certain locations, there was not a signigicant reduction in localisation performance. In general, the difference in performance between the simulated and measured radio maps was around a 30 % increase in rms error