液相色谱串联三重四极杆质谱(liqui dchromatography tandem triple quadrupole mass spectrometry,LC-MS/MS)广泛应用于环境分析、食品安全、药物代谢研究、生物医学等领域,主要用于复杂样品基质中痕量目标物检测,既适用于小分子有机...液相色谱串联三重四极杆质谱(liqui dchromatography tandem triple quadrupole mass spectrometry,LC-MS/MS)广泛应用于环境分析、食品安全、药物代谢研究、生物医学等领域,主要用于复杂样品基质中痕量目标物检测,既适用于小分子有机化合物定性,又可用于痕量化合物定量.其主要检测模式是多反应监测模式(multiple reaction monitoring,MRM),以定性离子对的丰度比来进行定性,以定量离子对的峰响应进行定量,在MRM检测模式下具有出色的检测灵敏度和定量功能.LC-MS/MS的合理规范使用与维护保养,是保证检测数据准确有效、仪器可持续正常运行的基本要求,也是降低仪器维修成本的重要举措.阐述了安捷伦LC-MS/MS的工作原理、开机与关机、调谐,探讨了常见的故障及应对措施、日常维护保养的内容,旨在为合理规范使用仪器提供参考.展开更多
[Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). [Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vect...[Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). [Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vector; the recombinants were digested by Nhe Ⅰ and Hind Ⅲ, and ligated into pEGFP-N1-P7.5, yielding the recombinant vector pEGFP-N1-P7.5-H; next the expression cassette EGFP-N1-P7.5-H was first released from recombinant vector pEGFP-N1-P7.5-H by double digestion of Hind Ⅲ and Nhe Ⅰ and ligated into pUC119-TK that was digested by Kpn Ⅰ, yielding the transfer vector pUC119-TK-EGFP-P7.5-H. [Result] Identification and double enzyme digestion showed that the transfer vector pUC119-TK-EGFP-P7.5-H was correctly constructed. From the transfer vector transfected BHK-21 cells which infected GTPV AV41, specific fluorescence was observed at 48th h of transfection. [Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine.展开更多
文摘液相色谱串联三重四极杆质谱(liqui dchromatography tandem triple quadrupole mass spectrometry,LC-MS/MS)广泛应用于环境分析、食品安全、药物代谢研究、生物医学等领域,主要用于复杂样品基质中痕量目标物检测,既适用于小分子有机化合物定性,又可用于痕量化合物定量.其主要检测模式是多反应监测模式(multiple reaction monitoring,MRM),以定性离子对的丰度比来进行定性,以定量离子对的峰响应进行定量,在MRM检测模式下具有出色的检测灵敏度和定量功能.LC-MS/MS的合理规范使用与维护保养,是保证检测数据准确有效、仪器可持续正常运行的基本要求,也是降低仪器维修成本的重要举措.阐述了安捷伦LC-MS/MS的工作原理、开机与关机、调谐,探讨了常见的故障及应对措施、日常维护保养的内容,旨在为合理规范使用仪器提供参考.
文摘[Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). [Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vector; the recombinants were digested by Nhe Ⅰ and Hind Ⅲ, and ligated into pEGFP-N1-P7.5, yielding the recombinant vector pEGFP-N1-P7.5-H; next the expression cassette EGFP-N1-P7.5-H was first released from recombinant vector pEGFP-N1-P7.5-H by double digestion of Hind Ⅲ and Nhe Ⅰ and ligated into pUC119-TK that was digested by Kpn Ⅰ, yielding the transfer vector pUC119-TK-EGFP-P7.5-H. [Result] Identification and double enzyme digestion showed that the transfer vector pUC119-TK-EGFP-P7.5-H was correctly constructed. From the transfer vector transfected BHK-21 cells which infected GTPV AV41, specific fluorescence was observed at 48th h of transfection. [Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine.
