Expression and purification of an adenylation domain from a eukaryotic nonribosomal peptide synthetase: Using structural genomics tools for a challenging target

Nonribosomal peptide synthetases (NRPSs) are large multimodular and multidomain enzymes that are involved in synthesising an array of molecules that are important in human and animal health. NRPSs are found in both bacteria and fungi but most of the research to date has focused on the bacterial enzymes. This is largely due to the technical challenges in producing active fungal NRPSs, which stem from their large size and multidomain nature. In order to target fungal NRPS domains for biochemical and structural characterisation, we tackled this challenge by using the cloning and expression tools of structural genomics to screen the many variables that can influence the expression and purification of proteins. Using these tools we have screened 32 constructs containing 16 different fungal NRPS domains or domain combinations for expression and solubility. Two of these yielded soluble protein with one, the third adenylation domain of the SidN NRPS (SidNA3) from the grass endophyte Neotyphodium lolii, being tractable for purification using Ni-affinity resin. The initial purified protein exhibited poor solution behaviour but optimisation of the expression construct and the buffer conditions used for purification, resulted in stable recombinant protein suitable for biochemical characterisation, crystallisation and structure determination

Fermions in 3D Optical Lattices: Cooling Protocol to Obtain Antiferromagnetism

A major challenge in realizing antiferromagnetic (AF) and superfluid phases in optical lattices is the ability to cool fermions. We determine the equation of state for the 3D repulsive Fermi-Hubbard model as a function of the chemical potential, temperature and repulsion using unbiased determinantal quantum Monte Carlo methods, and we then use the local density approximation to model a harmonic trap. We show that increasing repulsion leads to cooling, but only in a trap, due to the redistribution of entropy from the center to the metallic wings. Thus, even when the average entropy per particle is larger than that required for antiferromagnetism in the homogeneous system, the trap enables the formation of an AF Mott phase.Comment: 4 pages; 5 figures; also see supplementary material in 2 pages with 1 figur

Measurement of the Hyperfine Structure and Isotope Shifts of the 3s23p2 3P2 to 3s3p3 3Do3 Transition in Silicon

The hyperfine structure and isotope shifts of the 3s23p2 3P2 to 3s3p3 3Do3 transition in silicon have been measured. The transition at 221.7 nm was studied by laser induced fluorescence in an atomic Si beam. For 29Si, the hyperfine A constant for the 3s23p2 3P2 level was determined to be -160.1+-1.3 MHz (1 sigma error), and the A constant for the 3s3p3 3Do3 level is -532.9+-0.6 MHz. This is the first time that these constants were measured. The isotope shifts (relative to the abundant isotope 28Si) of the transition were determined to be 1753.3+-1.1 MHz for 29Si and 3359.9+-0.6 MHz for 30Si. This is an improvement by about two orders of magnitude over a previous measurement. From these results we are able to predict the hyperfine structure and isotope shift of the radioactive 31Si atom, which is of interest in building a scalable quantum computer

Stellar Populations and Star Cluster Formation in Interacting Galaxies with the Advanced Camera for Surveys

Pixel-by-pixel colour-magnitude and colour-colour diagrams - based on a subset of the Hubble Space Telescope Advanced Camera for Surveys Early Release Observations - provide a powerful technique to explore and deduce the star and star cluster formation histories of the Mice and the Tadpole interacting galaxies. In each interacting system we find some 40 bright young star clusters (20 <= F606W (mag) <= 25, with a characteristic mass of ~3 x 10^6 Msun), which are spatially coincident with blue regions of active star formation in their tidal tails and spiral arms. We estimate that the main events triggering the formation of these clusters occurred ~(1.5-2.0) x 10^8 yr ago. We show that star cluster formation is a major mode of star formation in galaxy interactions, with >= 35% of the active star formation in encounters occurring in star clusters. This is the first time that young star clusters have been detected along the tidal tails in interacting galaxies. The tidal tail of the Tadpole system is dominated by blue star forming regions, which occupy some 60% of the total area covered by the tail and contribute ~70% of the total flux in the F475W filter (decreasing to ~40% in F814W). The remaining pixels in the tail have colours consistent with those of the main disk. The tidally triggered burst of star formation in the Mice is of similar strength in both interacting galaxies, but it has affected only relatively small, spatially coherent areas.Comment: 23 pages in preprint form, 6 (encapsulated) postscript figures; accepted for publication in New Astronomy; ALL figures (even the grey-scale ones) need to be printed on a colour printer style files included; for full-resolution paper, see http://www.ast.cam.ac.uk/STELLARPOPS/ACSpaper

Next Generation Advanced Video Guidance Sensor Development and Test

The Advanced Video Guidance Sensor (AVGS) was the primary docking sensor for the Orbital Express mission. The sensor performed extremely well during the mission, and the technology has been proven on orbit in other flights too. Parts obsolescence issues prevented the construction of more AVGS units, so the next generation of sensor was designed with current parts and updated to support future programs. The Next Generation Advanced Video Guidance Sensor (NGAVGS) has been tested as a breadboard, two different brassboard units, and a prototype. The testing revealed further improvements that could be made and demonstrated capability beyond that ever demonstrated by the sensor on orbit. This paper presents some of the sensor history, parts obsolescence issues, radiation concerns, and software improvements to the NGAVGS. In addition, some of the testing and test results are presented. The NGAVGS has shown that it will meet the general requirements for any space proximity operations or docking need