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- Ding, HT, et al.
(författare)
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Parallel cloning, expression, purification and crystallization of human proteins for structural genomics
- 2002
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Ingår i: Acta Crystallographica. Section D: Biological Crystallography. - 1399-0047. ; 58, s. 2102-2108
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Tidskriftsartikel (refereegranskat)abstract
- 54 human genes were selected as test targets for parallel cloning, expression, purification and crystallization. Proteins from these genes were selected to have a molecular weight of between 14 and 50 kDa, not to have a high percentage of hydrophobic residues (i.e. more likely to be soluble) and to have no known crystal structures and were not known to be subunits of heterocomplexes. Four proteins containing transmembrane regions were selected for comparative tests. To date, 44 expression clones have been constructed with the Gateway(TM) cloning system (Invitrogen, The Netherlands). Of these, 35 clones were expressed as recombinant proteins in Escherichia coli strain BL21 (DE3)-pLysS, of which 12 were soluble and four have been purified to homogeneity. Crystallization conditions were screened for the purified proteins in 96-well plates under oil. After further refinement with the same device or by the hanging-drop method, crystals were grown, with needle, plate and prism shapes. A 2.12 Angstrom data set was collected for protein NCC27. The results provide insights into the high-throughput target selection, cloning, expression and crystallization of human genomic proteins.
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- Franz, P., et al.
(författare)
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Compact soft x-ray multichord camera : Design and initial operation
- 2003
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Ingår i: Review of Scientific Instruments. - : AIP Publishing. - 0034-6748 .- 1089-7623. ; 74:3 II, s. 2152-2156
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Tidskriftsartikel (refereegranskat)abstract
- The design and initial operation of a compact soft x-ray (SXR) multichord camera was studied. The line integrity emissivity was measured along up to 20 lines of sight using an array of miniaturized silicon photodiodes. The application of the diagnostic to a gas-fed magnetoplasma dynamic thruster (MPDT) with an external magnetic field was also discussed.
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- Rosén, Bengt-Göran, 1962-, et al.
(författare)
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Oil pockets and surface topography : Mechanisms of friction reduction
- 2004
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Oil pockets reduce friction in two ways: by providing lift themselves by a cavitation mechanism, and also by acting as a reservoir of lubricant which will leak out around their boundaries to minimise direct metallic contact in the surrounding region. The relative importance of these mechanisms is estimated in particular tribological environments. The general cavitation conditions for oil pockets are defined and the cavitation models of Etsion and Kato are compared. Both these models require knowledge of the size and distribution of oil pockets, and an attempt is made to relate these to measurable surface topography parameters. Mechanisms for leakage of lubricant from oil pockets are also discussed. Advantages and disadvantages of the Wihiborg-Crafoord index are presented. The effect of oil pockets on the contact regime in gears is assessed, and in particular the likely effect of oil-pocket-induced cavitation on gear noise, and its relation to existing work on gear roughness, is discussed. Problems of oil pocket measurement on reaÌ surfaces are outlined and the advantages of 3D measurement are identified.
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- Selmer, Maria, et al.
(författare)
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Preparation of a crystallizable mRNA-binding fragment of Moorella thermoacetica elongation factor SelB
- 2002
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Ingår i: Acta Crystallographica. Section D: Biological Crystallography. - 1399-0047. ; 58, s. 1871-1873
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Tidskriftsartikel (refereegranskat)abstract
- SelB is a bacterial elongation factor required for the decoding of a UGA stop codon together with a specific mRNA hairpin to selenocysteine. In attempts to crystallize Moorella thermoacetica SelB, a proteolysis process occurred and crystals of a proteolytic fragment were observed. The crystals, which appeared after a year, contained a C-terminal 30 kDa fragment containing the mRNA-binding domain. This fragment was reproduced through recloning. Crystals diffracting to 2.7 Angstrom were obtained.
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