Copurification of the Lac repressor with polyhistidine-tagged proteins in immobilized metal affinity chromatography.

One of the commonly used resins for immobilized metal affinity purification of polyhistidine-tagged recombinant proteins is TALON resin, a cobalt (II)--carboxymethylaspartate-based matrix linked to Sepharose CL-6B. Here, we show that TALON resin efficiently purifies the native form of Lac repressor, which represents the major contaminant when (His)(6)-tagged proteins are isolated from Escherichia coli host cells carrying the lacI(q) gene. Inspection of the crystal structure of the repressor suggests that three His residues (residues 163, 173, and 202) in each subunit of the tetramer are optimally spaced on an exposed face of the protein to allow interaction with Co(II). In addition to establishing a more efficient procedure for purification of the Lac repressor, these studies indicate that non-lacI(q)-based expression systems yield significantly purer preparations of recombinant polyhistidine-tagged proteins.

Vortex filament simulation of trailing vortex merger

Aircraft in high-lift configuration shed multiple vortices. These generally merge to form a downstream wake consisting of two counter-rotating vortices of equal strength. The understanding of the merger of two co-rotating trailing vortices is important in evaluating the separation criteria for different aircraft to prevent wake vortex hazards during landing and take-off. There is no existing theoretical method on the basis of which such norms can be set. The present study is aimed at gaining a better understanding of the behaviour of wake vortices behind the aircraft. Two dimensional studies are carried out using the vortex blob method and compared with Bertenyi's experiment. It is shown that inviscid two dimensional effects are insufficient to explain the observations. Three dimensional studies, using the vortex filament method, are applied to the same test case. Two Lamb-Oseen profile vortices of the same dimensions and initial separation as the experiment are allowed to evolve from a straight starting condition until a converged steady flow is achieved. The results obtained show good agreement with the experimental distance to merger. Core radius and separation behaviour is qualitatively similar to experiment, with the exception of rapid increases at first. This may be partially attributable to the choice of filament-element length, and recommended further work includes a convergence study for this parameter. Copyright © 2005 by the American Institute of Aeronautics and Astronautics, Inc. All rights reserved.

The oxidation of CH2O in the intermediate temperature range (943-995 K)

New atmospheric pressure flow reactor data on the oxidation of formaldehyde in the temperature range 943-995 K and over equivalence ratios from 0.013 to 36.7 are reported and discussed. A detailed mechanism assembled from previously published results produced acceptable agreement with the experimental data for the fuel-lean conditions, but failed to predict results for oxidative pyrolysis. Analysis or the very fuel-lean conditions, but failed to modelling results are principally sensitive to CH2O+HO2→HCO+H2O2 (6) and H2O2 +M→OH+OH+M (33). Whereas the specific rate of each reaction cannot be independently determined, it is found that the product k33.k6 is a well defined function of temperature: (3.4±3.0).1028 exp(-(26,800±400)/T). Inadequacies in the mechanism which may be responsible for the disagreement under fuel-rich conditions are discussed. © 1991 Combustion Institute.