[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan ...[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan three-way cross-breeding pigs. According to the porcine IL-18 gene in GenBank, a pair of specific primers was designed. The full length cDNA of porcine IL-18 was amplified by RT-PCR. Subsequently, porcine IL-18 cDNA was cloned into pGEM-T vector and sequenced and analyzed. [ Result] The porcine IL-18 gene demonstrated an open reading frame of 579 bp encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and cleaved by interleukin-1 beta （IL-1β） converting enzyme（ICE） in caspase-1 splice site; the porcine mature protein had biological activity： After comparing with other porcine IL-18 genes, the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [ Conclusion] A full length procine IL-18 gene was gained. It lays the foundation for porcine IL-18 as an adjuvant of genetic vaccine.展开更多
Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine...Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification （LAMP） assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase （empA） gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.展开更多
Detonation of low energy detonating fuse was studied in numerical simulation and experiments in bending conditions using LS_DYNA3D. The results show that pressure of the explosion and detonation velocity decrease in t...Detonation of low energy detonating fuse was studied in numerical simulation and experiments in bending conditions using LS_DYNA3D. The results show that pressure of the explosion and detonation velocity decrease in the same section areas after bending. In bending conditions, detonation wave was similar to small angle comer diffraction. So the detonation velocity was lower than normal velocity.展开更多
基金Supported by Project of Basic and Frontier Technology Research in Henan Province(072300430060 )Key Scientific and Technological Project of Henan Province(072102130023)~~
文摘[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan three-way cross-breeding pigs. According to the porcine IL-18 gene in GenBank, a pair of specific primers was designed. The full length cDNA of porcine IL-18 was amplified by RT-PCR. Subsequently, porcine IL-18 cDNA was cloned into pGEM-T vector and sequenced and analyzed. [ Result] The porcine IL-18 gene demonstrated an open reading frame of 579 bp encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and cleaved by interleukin-1 beta （IL-1β） converting enzyme（ICE） in caspase-1 splice site; the porcine mature protein had biological activity： After comparing with other porcine IL-18 genes, the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [ Conclusion] A full length procine IL-18 gene was gained. It lays the foundation for porcine IL-18 as an adjuvant of genetic vaccine.
基金Supported by the National Basic Research Program of China (973 Program) (No. 2006CB101804)the General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China (No. 2007IK167)National Department Public Benefit Research Foundation (No. 200803012)
文摘Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming. The rapid detection and identification of V. anguillarum, and other pathogens that infect marine organisms, is crucial to effective disease management. In this study, we developed a loop-mediated amplification （LAMP） assay to detect V. anguillarum in an hour in a single tube without the need for thermal cycling. Conserved regions of the metalloproteinase （empA） gene of V. anguillarum served as the targets for primer design. A fragment of the empA gene was amplified at 65℃ in the presence of the primer mixture and Bst DNA polymerase. In the optimized LAMP assay, 6.7 pg of V. anguillarum DNA could be detected. Six strains of V. anguillarum and 17 strains of non-V, anguillarum bacteria were used in this study to evaluate the species specificity of the primers. The six V. anguillarum strains gave a positive result in the LAMP assay. This method was also validated in V. anguillarum-infected fish. This LAMP method is more sensitive than PCR in the detection of V. anguillarum and shows good species specificity. The LAMP assay is therefore an effective method for the quick detection of V. anguillarum both in the laboratory and in the field.
文摘Detonation of low energy detonating fuse was studied in numerical simulation and experiments in bending conditions using LS_DYNA3D. The results show that pressure of the explosion and detonation velocity decrease in the same section areas after bending. In bending conditions, detonation wave was similar to small angle comer diffraction. So the detonation velocity was lower than normal velocity.
