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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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| 3. |
- Li, Z-F, et al.
(författare)
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Determination of carrier-transfer length from side-wall quantum well to quantum wire by micro-photoluminescence scanning
- 2003
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Ingår i: Journal of Electronic Materials. - : Springer Science Business Media. - 0361-5235 .- 1543-186X. ; 32:8, s. 913-916
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Tidskriftsartikel (refereegranskat)abstract
- Micro-photoluminescence (mu-PL) line scanning across a single V-groove, GaAs/AlGaAs quantum wire (QWR) has been performed at room temperature, revealing a clear spatial-dependence of the PL. After fitting each PL spectrum by multi-Gaussian line shapes, intensity profiles of each PL component from confined structures have been obtained as functions of the scanning position. The PL quenching of a side-wall quantum well (SQWL) has been recognized in a certain area in the vicinity of the QWR and is interpreted by carrier transfer into the QWR within effective transfer length. By simulating the carrier-transfer process from SQWL to QWR as a convolution of a step function for carrier distribution and a Gaussian function for exciting laser irradiance, the effective transfer length of about 1.8+/-0.3 mum has, therefore, been concluded.
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| 7. |
- Scherer, SW, et al.
(författare)
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Human chromosome 7: DNA sequence and biology
- 2003
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Ingår i: Science (New York, N.Y.). - : American Association for the Advancement of Science (AAAS). - 1095-9203 .- 0036-8075. ; 300:5620, s. 767-772
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Tidskriftsartikel (refereegranskat)abstract
- DNA sequence and annotation of the entire human chromosome 7, encompassing nearly 158 million nucleotides of DNA and 1917 gene structures, are presented. To generate a higher order description, additional structural features such as imprinted genes, fragile sites, and segmental duplications were integrated at the level of the DNA sequence with medical genetic data, including 440 chromosome rearrangement breakpoints associated with disease. This approach enabled the discovery of candidate genes for developmental diseases including autism.
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