Eukaryotic expression, purification and activity characterization of human soluble DSG2 extracellular domain protein

Objective:To construct a secretory eukaryotic expression vector of DSG2 fused with the Fc region of the human IgG,to validate its expression in 293T cells,and to purify the secretory protein with biological activity.Methods:The DSG2 extracellular domain fragment gene(DSG2ex),was amplified by PCR,and was inserted into the eukaryotic expression plasmid pCMV3-IgG1 to construct the recombinant eukaryotic expression plasmid-pCMV3-DSG2ex-IgG1.The successfully constructed eukaryotic expression plasmid was transfected into 293T cells to express and secrete DSG2 extracellular domain protein.The targeted protein was purified from the cell culture supernatant by Protein A affinity chromatography and confirmed by Western Blotting and ELISA.Results:The pCMV3-DSG2ex-IgG1 eukaryotic expression plasmid was successfully constructed.The highest protein expression level was obtained with 293T cells after 96 h of transfection.The relative molecular mass of the purified product was between 100 and 130 kDa was estimated by SDS-PAGE,which was consistent with the expectation.The yield of the purified protein reached 0.8 mg/ml with a purity over 90%.The purified DSG2 extracellular domain protein with IgG1 tag was recognized by IgG monoclonal antibodies by Western blotting.Moreover,the ELISA results showed that the prepared DSG2 extracellular domain protein had significant binding activity to human type 55 adenovirus Fiber Knob protein(HAdV-55).Conclusion:A simple and efficient method for eukaryotic expression and purification of human soluble DSG2 extracellular domain protein was successfully established,and biologically active DSG2 extracellular domain protein was purified,which laid the foundation for the later study of its protein function and anti-adenovirus drugs.

Characterization and Methanol Adsorption of Walnutshell Activated Carbon Prepared by KOH Activation

Walnut-shellactivated carbons（WSACs）were prepared by the KOH chemicalactivation.The effects of carbonization temperature,activation temperature,and ratio of KOH to chars on the pore development of WSACs were investigated.Fourier transform infrared spectroscopy（FTIR）,X-ray powder diffraction（XRD）,and scanning electron microscopy（SEM）were employed to characterize the microstructure and morphology of WSACs.Methanoladsorption performance onto the optimalWSAC and the coal-based AC were also investigated.The results show that the optimalpreparation conditions are a carbonization temperature of 700 ℃,an activation temperature of 700 ℃,and a mass ratio of 3.The BET surface area,the micropore volume,and the micropore volume percentage of the optimalWASC are 1636 m^2/g,0.641 cm^3/g and 81.97%,respectively.There are a lot of micropores and a certain amount of meso-and macropores.The characteristics of the amorphous state are identified.The results show that the optimalWSAC is favorable for methanoladsorption.The equilibrium adsorption capacity of the optimalWSAC is 248.02mg/g.It is shown that the equilibrium adsorption capacity of the optimalWSAC is almost equivalent to that of the common activated carbon.Therefore the optimalWSAC could be a potentialadsorbent for the solar energy adsorption refrigeration cycle.

Fuzzy statistic and comprehensive evaluating study for activity characterization of the active region

In this paper, the theory and method of the fuzzy mathematics areused to probe the connection between the activity of the active region and characterizat ion of the sunspot groups, to build the subordinating function according to the rela tionship between them and to evaluate comprehensively the activity of the active region on t he solar disk. The precise prediction of activity of the active regions has been obta ined by data reduction and analysis. The predicting accuracy is higher th an 95% . Forecast results indicate that the method of the fuzzy comprehensive evaluatio n is a good one for the solar activity prediction.

Characterization of geometry and depleting carrier dependence of active silicon waveguide in tailoring optical properties

Changes in refractive index and the corresponding changes in the characteristics of an optical waveguide in enabling propagation of light are the basis for many modern silicon photonic devices. Optical properties of these active nanoscale waveguides are sensitive to the little changes in geometry, external injection/biasing, and doping profiles, and can be crucial in design and manufacturing processes. This paper brings the active silicon waveguide for complete characterization of various distinctive guiding parameters, including perturbation in real and imaginary refractive index, mode loss, group velocity dispersion, and bending loss, which can be instrumental in developing optimal design specifications for various application-centric active silicon waveguides.

Synthesis and spectroscopic characterization of cobalt porphyrins and their catalytic activity towards dioxygen reduction

The synthesis and spectroscopic characterization of cobalt(Ⅱ) 5-(4-pyridyl)-10,15,20-triphe- nylporphyrin,cobalt(Ⅱ) 5-(4-N-hexadecylpyridiniumyl)-10,15,20-triphenylporphyrin bromide and cobalt(Ⅱ) 5-(2-aminophenyl)-10,15,20-triphenyl-porphyrin are reported.The corresponding copper and vanadyl derivatives ((TriP)Cu,[(hTriP)Cu]^+Br^- and [(hTriP)VO]^+Br^-) were also studied.Each metalloporphyrin was characterized by UV-visible,ESR and ~1H NMR spectroscopy.These me- talloporphyrins can be firmly adsorbed on the glassy carbon (GC) surface.The catalytic reduction of dioxygen at GC electrodes modified by these catalysts was studied by cyclic voltammetry (CV).The kinetic process of dioxygen reduction at the cobalt porphyrin-modified electrodes was studied with a rotating ring disk electrode.

Carbon nanotubes/TiO_2 nanotubes composite photocatalysts for efficient degradation of methyl orange dye

A series of carbon nanotubes/TiO2 nanotubes （CNTs/TNTs） composite photocatalysts were successfully prepared by incorporation of CNTs in HNO3 washing process. These photocatalysts were characterized by XRD, N2 physical adsorption, UV-vis diffuse reflectance spectroscopy, TEM and Raman spectroscopy, respectively, and their photocatalytic activities were tested by using methyl orange （MO） as a model compound. Also, the effects of amount of CNTs incorporated, calcination temperature and amount of catalyst on the photocatalytic activity of the composite photocatalyst were systematically investigated. The results show that the CNTs/TNTs composite exhibits much higher photocatalytic activity than that of the TNTs or CNTs alone.