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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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- Zhang, Shi-Li, et al.
(författare)
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Abnormal anti-Stokes Raman scattering of carbon nanotubes
- 2002
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Ingår i: Physical Review B. Condensed Matter and Materials Physics. - 1098-0121 .- 1550-235X. ; 66:3
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Tidskriftsartikel (refereegranskat)abstract
- Abnormal anti-Stokes Raman scattering (AASR) was unambiguously observed in carbon nanotubes (CNT's). In contrast to traditional Raman scattering theory, the absolute value of the Raman frequency of the anti-Stokes peak is not the same as that of the corresponding Stokes peak. It was demonstrated that AASR scattering originates from the unique nanoscale cylindrical structure of CNT's that can be considered naturally as a graphite structure with an intrinsic defect from its rolling. The double-resonance Raman scattering theory was applied to interpret the scattering mechanism of the AASR phenomenon successfully and quantitatively.
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| 13. |
- Hu, Hanmei, et al.
(författare)
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Solvothermal synthesis of Sb2S3 nanowires on a large scale
- 2003
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Ingår i: Journal of Crystal Growth. - : Elsevier. - 0022-0248 .- 1873-5002. ; 258:1-2, s. 106-112
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Tidskriftsartikel (refereegranskat)abstract
- Sb2S3 nanowires with high aspect ratios have been successfully prepared on a large scale using SbCl3 and Na2S as starting materials in ethylene glycol at 200°C for 10 h. Field emission scanning electron microscopy images and transmission electron microscopy images show that the nanowires have diameters in the range of 20–100 nm and lengths up to 50 μm. Diffuse reflection spectrum indicates that the as-prepared Sb2S3 nanowires have obvious quantum size effects. The effects of reaction parameters on the growth of nanowires were discussed. A possible mechanism on the formation of the Sb2S3 nanowires was proposed.
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| 20. |
- Li, Zhijun, et al.
(författare)
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Product tolerance allocation in compliant multistation assembly through variation propagation and analytical target cascading
- 2004
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Ingår i: Proceedings of the ASME Design Engineering Division - 2004. - New York : American Society of Mechanical Engineers. - 0791847055 ; , s. 813-820
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Konferensbidrag (refereegranskat)abstract
- Compliant sheet metal assembly is a hierarchical manufacturing process that plays a significant role in automotive product development. Parts are joined in different stations to form the final product (e.g., the vehicle body structure). Dimensional variation is a product attribute of major importance that characterizes quality, and is mainly affected by the variability of parts, fixtures, and joining methods at each of the multiple stations. The propagation of dimensional variation through the multistation assembly system is modeled as a linear process, where all three aforementioned sources of variability are taken into account at each station using finite element models. In this article we apply the analytical target cascading process to the tolerance allocation problem in multistation assembly systems. Specifically, we translate final product variation targets to tolerance specifications for subassemblies and incoming parts. We demonstrate the methodology by means of a vehicle side frame assembly example.
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- Zhuang, H, et al.
(författare)
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Cloning of a T-type Ca2+ channel isoform in insulin-secreting cells
- 2000
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Ingår i: Diabetes. - : American Diabetes Association. - 0012-1797 .- 1939-327X. ; 49:1, s. 59-64
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Tidskriftsartikel (refereegranskat)abstract
- The T-type Ca2+ channel is an important determinant of electrical activity and of Ca2+ influx in rat and human pancreatic beta-cells. We have identified and sequenced a cDNA encoding a T-type Ca2+ channel alpha1-subunit derived from INS-1, the rat insulin-secreting cell line. The sequence of the cDNA indicates a protein composed of 2,288 amino acids that shares 96.3% identity to alpha1G, the neuronal T-type Ca2+ channel subunit. The transmembrane domains of the protein are highly conserved, but the isoform contains three distinct regions and 10 single amino acid substitutions in other regions. Sequencing rat genomic DNA revealed that the alpha1-subunit we cloned is an alternative splice isoform of alpha1G. By using specific primers and reverse transcription-polymerase chain reaction, we demonstrated that both splice variants are expressed in rat islets. The isoform deduced from INS-1 was also expressed in brain, neonatal heart, and kidney. Functional expression of this alpha1G isoform in Xenopus oocytes generated low voltage-activated Ba2+ currents. These results provide the molecular biological basis for studies of function of T-type Ca2+ channels in beta-cells, which is where these channels may play critical roles in diabetes.
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