Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli

Aerolysin is a toxin (protein in nature) secreted by the strains of Aeromonas spp. and plays all important role in the virulence of Aeromonas strains. It has also found several applications such as for detection of glycosylphosphatidylinositol (GPI)-anchored proteins etc. A. hydrophila is a ubiquitous Gram-negative bacterium which causes frequent harm to the aquaculture. To obtain a significant amount of recombinant aerolysin in the active form, in this study, we expressed the aerolysin in E. Coli Under the control of T7 RNase promoter. The coding region (AerA-W) of the aerA gene of A. hydrophila XS91-4-1. excluding partial coding region of the signal peptide was cloned into the vector pET32a and then transformed into E. coli b121. After optimizing the expression conditions, the recombinant protein AerA-W was expressed in a soluble form and purified using His-Bind resin affinity chromatography. Recombinant aerolysin showed hemolytic activity in the agar diffusive hemolysis test. Western blot analysis demonstrated good antigenicity of the recombinant protein.

Purification and characterization of recombinant truncated human interleukin-11 expressed as fusion protein in Escherichia coli

Mature human interleukin-11 (HuIL-11) is a cytokine consisting of 178 amino acid residues that results from scission of the N-terminal signal peptide, consisting of 21 amino acid residaues, from the corresponding nascent polypeptide. A DNA fragment encoding a truncated HuIL-11 (trHuIL-11), with an additional 5 amino acid residues removed from the N-terminus, was cloned into vector pGEX-2T between the BamHI site and the EcoRI site. Upon transformation with Escherichia coli BL21, the construct over-produced a glutathione S-transferase (GST)-fused protein in a soluble form after IPTG induction. The fusion protein was initially fractionated with butyl-Sepharose 4 fast flow column and by affinity chromatography using a GSH-Sepharose 4B column. On-site enzymatic release with thrombin gave the target protein at 96% purity as judged by SDS-PAGE and HPLC. Expression of the interleukin as a GST-fused protein thus greatly improved downstream processing. Subsequent biological activity assay suggested that trHuIL-11 had similar activity profile to the naturally produced sample and may be a promising candidate for further development as biopharmaceutical.

Five supramolecular compounds of water-soluble sulfonylcalix[4]arenetetrasulfonate showing two calixarene conformations

Five new compounds of sulfonylcalix[4]arenetetrasulfonate (SC4AS), [H7Na(H2O)(3)(SC4AS)(phen)(5)](H2O)(11.9) (1), [H6Mn(H2O)(4)(SC4AS)(phen)(5)] (H2O)(12.7) (2), [Cu-4(SC4AS) (phen)(6)] (H2O)(4.5) (3), {[Cu (2)(SC4AS) (bpy)(2)][Cu(bpy)(2)(H2O)](2)} (H2O)(6.6) (4), and {[Zn-2(SC4AS) (phen)(2)][Zn(phen)(2)(H2O)(2)](2)} (H2O)(7) (5) (where phen 1,10-phenanthroline and bpy = 2,2'-bipyridine), were synthesized by a hydrothermal method and structurally determined by single crystal X-ray diffraction. The SC4AS ligand adopts partial cone conformation in compounds 1 and 2 and 1,2-alternate form in compounds 3-5. According to the structural analysis and density functional theory (DFT) calculations, we suggest that the metal can affect the conformation of SC4AS.

Closed form solutions of T-stress in plane elasticity crack problems

The T-stress is considered as an important parameter in linear elastic fracture mechanics. In this paper, several closed form solutions of T-stress in plane elasticity crack problems in an infinite plate are investigated using the complex potential theory. In the line crack case, if the applied loading is the remote stress or the concentrated forces, the T-stress can be derived from the basic field. Here, the basic field is defined as the field caused by the applied loading in the infinite plate without the crack. For the circular are crack, the T-stress can be abstracted from a known solution. For the cusp crack problems, the T-stress can be separated from the obtained stress solution for which the conformal mapping technique is used.

Soluble Angiopoietin Receptor Tie-2 In Patients With Acute Myocardial Infarction And Its Detection By Optical Protein-Chip

The Tie-2 receptor has been shown to play a role in angiogenesis in atherosclerosis. The conventional method assaying the level of soluble Tie-2 (sTie-2) was ELISA. However, this method has some disadvantages. The aims of this research are to establish a more simple detection method, the optical protein-chip based on imaging ellipsomtry (OPC-IE) applying to Tie-2 assay. The sTie-2 biosensor surface on silicon wafer was prepared first, and then serum levels of sTie-2 in 38 patients with AMI were measured on admission (day 1), day 2, day 3 and day 7 after onset of chest pain and 41 healthy controls by ELISA and OPC-IE in parallel. Median level of sTie-2 increased significantly in the AMI patients when compared with the controls. Statistics showed there was a significant correlation in sTie-2 results between the two methods (r=0.923, P0.01). The result of this study showed that the level of sTie-2 increased in AMI, and OPC-IE assay was a fast, reliable, and convenient technique to measure sTie-2 in serum.

Precipitation of nanocrystals in glasses by electron irradiation: An alternative path to form glass ceramics?

This letter demonstrates an alternative method to form gallium silicate glass ceramics using high-energy electron irradiation. Compared with glass ceramics obtained from the conventional thermal treatment method, the distribution and crystal sizes of the precipitated Ga2O3 nanoparticles are the same. An advantage of this method is that the spatial distribution of the precipitated nanoparticles can be easily controlled. However, optically active dopants Ni2+ ions do not participate in the precipitation during electron irradiation. (c) 2007 American Institute of Physics.

Preparation, characterization and gas sensing properties of high soluble metal (II) phthalocyanine thin films by spin-coating method

Three novel metal (II) phthalocyanine complexes were synthesized by cyclic tetramerisation reaction of a dicyano benzene component and different metal ions (Pd2+, Co2+, Zn2+). The structure of complexes was confirmed by elemental analysis, mass and IR spectrum. The excellent solubility of the complexes in benzene enabled us to obtain films by a spin-coating method. The films were characterized by IR, electronic spectral and AFM. The gas sensing properties to NO2 of the metal (II) phthalocyanine complex films were studied. In addition, the effects of different metal ions and the gas sensing temperature on the sensing properties were studied. (C) 2005 Elsevier B.V. All rights reserved.

Form Birefringence in thin films with oblique columnar structures

Effective medium theory is useful for designing optical elements with form birefringent subwavelength structures. Thin films fabricated by oblique deposition are similar to the two-dimensional surface relief subwavelength gratings. We use the effective medium theory to calculate the anisotropic optical properties of the thin films with oblique columnar structures. The effective refractive indices and the directions are calculated from effective medium theory. It is shown that optical thin films with predetermined refractive indices and birefringence may be engineered.