AIM: To compare the efficiency of different plasmids as DNA vectors by cloning three HBsAg-encoded genes into two eukaryotic expression vectors, pRc/CMV and pSG5UTPL/Flag, and to express HBsAg S, MS, and LS proteins i...AIM: To compare the efficiency of different plasmids as DNA vectors by cloning three HBsAg-encoded genes into two eukaryotic expression vectors, pRc/CMV and pSG5UTPL/Flag, and to express HBsAg S, MS, and LS proteins in SP2/0 cells, and to establish monoclone SP2/0 cell strains that are capable of expressing S or S2S proteins stably.METHODS: Segments of S, preS2-S, preS1-preS2-S genes of Hepatitis B virus were amplified by routine PCR and preS1S fragment was amplified by Over-Lap Extension PCR. The amplified segments were cleaved with restricted endonuclease Hind Ⅲ/Not Ⅰ followed by ligation with pRc/CMV, or BamHI/EcoR Ⅰ followed by ligation with pSG5UTPL/Flag. After the plasmid vectors were cleaved with the correspond enzymes, the amplified segments were inserted into pRc/CMV or pSGSUTPL/Flag plasmid vectors with T4DNA ligase. KOZAK sequence was added before the initial ATG code of each fragment using specific primer. The inserted segments in the recombinant plasmids were sequenced after subcloning. BALB/c mice myeloma cells (SP2/0 cell line) were transfected with the recombinant plasmids. The expressions of the different recombinants were compared by Western-blot, using a monoclonal anti-HBs antibody as the primary antibody and peroxidase-labeled multi-linker as the secondary. Stable SP2/0-pRc/CMV-S or SP2/0- pRc/CMV-MS clones were established through clone screening with G418.RESULTS: Fragments with anticipated size were harvested after PCR. After recombination and screening, the sequences of the inserted segments in the recombinants were confirmed to be S, preS2S, preSl-preS2S and preSlS encoding genes,determined by sequencing. The results of Western-blot hybridization were positive for the anticipated proteins.Among them, pRc/CMV-S or pRc/CMV-MS demonstrated the highest expressing their respective antigen.CONCLUSION: Eight recombinant plasmids expressing S,M, L or preSlS proteins are obtained. For hepatitis surface antigen expression in eukaryotic cells, the vector pRc/CMV is superior to pSG5UTPL/Flag, and pRc/CMV-S and pRc/CMVMS are the most efficient in the pRc/CMV clones. SP2/0 cells stably expressing HBsAg are established, and may be used as target cells for evaluating the CTL activity of a DNA vaccine in vitro.展开更多
A simple, sensitive and convenient ion chromatography(IC) method was established for the simultaneous determination of twelve water-soluble inorganic anions(F -, Cl -, NO - 2, NO - 3, SO 2- 3, SO 2- 4, PO 3- 4), and f...A simple, sensitive and convenient ion chromatography(IC) method was established for the simultaneous determination of twelve water-soluble inorganic anions(F -, Cl -, NO - 2, NO - 3, SO 2- 3, SO 2- 4, PO 3- 4), and fifteen water-soluble organic ions(formate, acetate, MSA, oxalate, malonate, succinate, phthalates, etc.) in atmospheric aerosols. The linear concentrations ranged from 0.005 μg/m 3 to 500 μg/m 3(r = 0.999—0.9999). The relative standard deviation(RSD) were 0.43%—2.00% and the detection limits were from 2.7 ng/m 3 to 88 ng/m 3. The proposed method was successfully applied to the simultaneous determination of those inorganic ions and organic ions in PM 2.5 of Beijing.展开更多
Two new blue luminescent zinc and beryllium complexes with Schiff base calixarene derivative as the ligand were prepared. Their luminescent properties were determined, which indicated that they had strong blue fluores...Two new blue luminescent zinc and beryllium complexes with Schiff base calixarene derivative as the ligand were prepared. Their luminescent properties were determined, which indicated that they had strong blue fluorescent properties. They also had good solubility and film formation. These new complexes can be used as blue organic electroluminescent materials (OELMs) in organic electroluminescent devices.展开更多
We report the excited-state upconversion in Pr(0.5)Yb(1.5):ZBLAN under two-colour excitation of 960nm laser and xenon lamp light. Three obvious upconversion excitation peaks around 856.0, 804.2 and 787.1 nm were obser...We report the excited-state upconversion in Pr(0.5)Yb(1.5):ZBLAN under two-colour excitation of 960nm laser and xenon lamp light. Three obvious upconversion excitation peaks around 856.0, 804.2 and 787.1 nm were observed and the involved mechanism has been explained. The measured upconversion excitation peak 856.0nm corresponds to the sum of theoretical values 852nm and 866nm owing to the ^1G4(Pr^3+)→^1I6(Pr^3+) and ^1G4(Pr^3+)→^3P1(Pr^3+) excited state absorption transitions. The measured 804.2 and 787.1 nm upconversion excitation peaks originate from the excited state absorption transitions ^3H6(Pr^3+)→^1D2(Pr^3+) and ^1G4(Pr^3+)→^3P2(Pr^3+), respectively. The excited state absorption upconversion ^1G4(Pr^3+)→^1I6(Pr^3+) is strong because of its large oscillator strength f=23.040×10^-6.展开更多
N-pivot lariat ethers with picrylamino group as a chromophore (1, 2 and 3) have been prepared by reaction of N-(4-aminoaryl)monoaza crown ethers with picryl chrolide, and the selective coloration of 1, 2 and 3 for alk...N-pivot lariat ethers with picrylamino group as a chromophore (1, 2 and 3) have been prepared by reaction of N-(4-aminoaryl)monoaza crown ethers with picryl chrolide, and the selective coloration of 1, 2 and 3 for alkali metal salts and amines has been studied by UV-Vis spectra.展开更多
A novel fluorene-based conjugated copolymer containing cyclobutenedione unit was synthesized by Suzuki reaction.Its structure and properties were characterized by FTIR,^1HNMR,elemental analysis,PL spectroscopy,DSC,TGA...A novel fluorene-based conjugated copolymer containing cyclobutenedione unit was synthesized by Suzuki reaction.Its structure and properties were characterized by FTIR,^1HNMR,elemental analysis,PL spectroscopy,DSC,TGA and cyclic voltammetry.The resulting polymer shows strong yellow PL emission(561nm)and good solubility in polar aprotic solvents,i.e.THF,DMF,DMAC,DMSO,etc.DSC and TGA studies reveal that the novel polymer possesses excellent thermal stability with high glass transition temperature of 127℃ and onset decomposition temperature of 411℃.Cyclic voltammetry measurement demonstrated that the polymer has both hole and electron-transporting property.展开更多
基金the National Science Foundation of China,No. 39670670
文摘AIM: To compare the efficiency of different plasmids as DNA vectors by cloning three HBsAg-encoded genes into two eukaryotic expression vectors, pRc/CMV and pSG5UTPL/Flag, and to express HBsAg S, MS, and LS proteins in SP2/0 cells, and to establish monoclone SP2/0 cell strains that are capable of expressing S or S2S proteins stably.METHODS: Segments of S, preS2-S, preS1-preS2-S genes of Hepatitis B virus were amplified by routine PCR and preS1S fragment was amplified by Over-Lap Extension PCR. The amplified segments were cleaved with restricted endonuclease Hind Ⅲ/Not Ⅰ followed by ligation with pRc/CMV, or BamHI/EcoR Ⅰ followed by ligation with pSG5UTPL/Flag. After the plasmid vectors were cleaved with the correspond enzymes, the amplified segments were inserted into pRc/CMV or pSGSUTPL/Flag plasmid vectors with T4DNA ligase. KOZAK sequence was added before the initial ATG code of each fragment using specific primer. The inserted segments in the recombinant plasmids were sequenced after subcloning. BALB/c mice myeloma cells (SP2/0 cell line) were transfected with the recombinant plasmids. The expressions of the different recombinants were compared by Western-blot, using a monoclonal anti-HBs antibody as the primary antibody and peroxidase-labeled multi-linker as the secondary. Stable SP2/0-pRc/CMV-S or SP2/0- pRc/CMV-MS clones were established through clone screening with G418.RESULTS: Fragments with anticipated size were harvested after PCR. After recombination and screening, the sequences of the inserted segments in the recombinants were confirmed to be S, preS2S, preSl-preS2S and preSlS encoding genes,determined by sequencing. The results of Western-blot hybridization were positive for the anticipated proteins.Among them, pRc/CMV-S or pRc/CMV-MS demonstrated the highest expressing their respective antigen.CONCLUSION: Eight recombinant plasmids expressing S,M, L or preSlS proteins are obtained. For hepatitis surface antigen expression in eukaryotic cells, the vector pRc/CMV is superior to pSG5UTPL/Flag, and pRc/CMV-S and pRc/CMVMS are the most efficient in the pRc/CMV clones. SP2/0 cells stably expressing HBsAg are established, and may be used as target cells for evaluating the CTL activity of a DNA vaccine in vitro.
文摘A simple, sensitive and convenient ion chromatography(IC) method was established for the simultaneous determination of twelve water-soluble inorganic anions(F -, Cl -, NO - 2, NO - 3, SO 2- 3, SO 2- 4, PO 3- 4), and fifteen water-soluble organic ions(formate, acetate, MSA, oxalate, malonate, succinate, phthalates, etc.) in atmospheric aerosols. The linear concentrations ranged from 0.005 μg/m 3 to 500 μg/m 3(r = 0.999—0.9999). The relative standard deviation(RSD) were 0.43%—2.00% and the detection limits were from 2.7 ng/m 3 to 88 ng/m 3. The proposed method was successfully applied to the simultaneous determination of those inorganic ions and organic ions in PM 2.5 of Beijing.
基金Project 29972032 was supported by the National Natural Science Foundation of China.
文摘Two new blue luminescent zinc and beryllium complexes with Schiff base calixarene derivative as the ligand were prepared. Their luminescent properties were determined, which indicated that they had strong blue fluorescent properties. They also had good solubility and film formation. These new complexes can be used as blue organic electroluminescent materials (OELMs) in organic electroluminescent devices.
文摘We report the excited-state upconversion in Pr(0.5)Yb(1.5):ZBLAN under two-colour excitation of 960nm laser and xenon lamp light. Three obvious upconversion excitation peaks around 856.0, 804.2 and 787.1 nm were observed and the involved mechanism has been explained. The measured upconversion excitation peak 856.0nm corresponds to the sum of theoretical values 852nm and 866nm owing to the ^1G4(Pr^3+)→^1I6(Pr^3+) and ^1G4(Pr^3+)→^3P1(Pr^3+) excited state absorption transitions. The measured 804.2 and 787.1 nm upconversion excitation peaks originate from the excited state absorption transitions ^3H6(Pr^3+)→^1D2(Pr^3+) and ^1G4(Pr^3+)→^3P2(Pr^3+), respectively. The excited state absorption upconversion ^1G4(Pr^3+)→^1I6(Pr^3+) is strong because of its large oscillator strength f=23.040×10^-6.
文摘N-pivot lariat ethers with picrylamino group as a chromophore (1, 2 and 3) have been prepared by reaction of N-(4-aminoaryl)monoaza crown ethers with picryl chrolide, and the selective coloration of 1, 2 and 3 for alkali metal salts and amines has been studied by UV-Vis spectra.
文摘A novel fluorene-based conjugated copolymer containing cyclobutenedione unit was synthesized by Suzuki reaction.Its structure and properties were characterized by FTIR,^1HNMR,elemental analysis,PL spectroscopy,DSC,TGA and cyclic voltammetry.The resulting polymer shows strong yellow PL emission(561nm)and good solubility in polar aprotic solvents,i.e.THF,DMF,DMAC,DMSO,etc.DSC and TGA studies reveal that the novel polymer possesses excellent thermal stability with high glass transition temperature of 127℃ and onset decomposition temperature of 411℃.Cyclic voltammetry measurement demonstrated that the polymer has both hole and electron-transporting property.
