Phosphomannose isomerase （PMI） encoding gene manA is a desirable selective marker in transgenic research. Under- standing of its expression patterns in transgenic plant and establishing highly sensitive detection me...Phosphomannose isomerase （PMI） encoding gene manA is a desirable selective marker in transgenic research. Under- standing of its expression patterns in transgenic plant and establishing highly sensitive detection method based on immunoassay have great impacts on the application of PMI. In this study, PMI-specific monoclonal antibodies were generated using recombinant protein as immunogen, and could be used in Western blot to detect as little as 0.5 ng His-tagged PMI protein or rice expressed PMI protein in sample accounted for 0.4% of single rice grain （about 0.08 mg）. PMI protein driven by CaMV-35S promoter was detected in dozens of tested tissues, including root, stem, leaf, panicle, and seed at all developmental stages during rice growing, and PMI protein accounted for about 0.036% of total protein in the leaves at seedling stage. The established method potentially can be used to monitor PMI protein in rice grains.展开更多
目的分析张家港市2018年检出的65株沙门菌的血清型特征及分布特点,探讨该地区沙门菌优势菌群的变迁。方法收集张家港市2018年来自体检、食物中毒、食源性疾病主动监测等人源性沙门菌65株,运用脉冲场凝胶电泳(Pulsed Field Gel Electroph...目的分析张家港市2018年检出的65株沙门菌的血清型特征及分布特点,探讨该地区沙门菌优势菌群的变迁。方法收集张家港市2018年来自体检、食物中毒、食源性疾病主动监测等人源性沙门菌65株,运用脉冲场凝胶电泳(Pulsed Field Gel Electrophoresis,PFGE)技术对其中优势血清群进行分子分型,利用PulseNet China网络获得相关性指纹图谱并进行聚类分析。结果65株沙门菌分离株分布6个血清群和9个血清型,以D群为主(46.97%)、B群次之(21.21%)。31株D群沙门菌分为6个基因型(相似值100.00%为同一PFGE型别),条带相似度为71.63%~100.00%,2个克隆系(相似度为0.85~1.00的为同一PFGE的克隆系),2个克隆系T1与T2相似度为71.63%,克隆系相似度较高。14株B群沙门菌PFGE指纹图谱可分为12个克隆系13种基因型,条带相似度为10.21%~100.00%。结论张家港市2018年人源沙门菌血清型呈多样化分布,优势血清群D群PFGE带型存在较近亲缘关系,B群沙门菌PFGE带型较多,基因多态性较高。展开更多
基金supported in part by the Natural Science Foundation of Beijing, China (5121001)the Cultivate New Varieties of Genetically Modified Organisms Technology Major Projects, the Ministry of Science and Technology of China (2009ZX08012-006B)
文摘Phosphomannose isomerase （PMI） encoding gene manA is a desirable selective marker in transgenic research. Under- standing of its expression patterns in transgenic plant and establishing highly sensitive detection method based on immunoassay have great impacts on the application of PMI. In this study, PMI-specific monoclonal antibodies were generated using recombinant protein as immunogen, and could be used in Western blot to detect as little as 0.5 ng His-tagged PMI protein or rice expressed PMI protein in sample accounted for 0.4% of single rice grain （about 0.08 mg）. PMI protein driven by CaMV-35S promoter was detected in dozens of tested tissues, including root, stem, leaf, panicle, and seed at all developmental stages during rice growing, and PMI protein accounted for about 0.036% of total protein in the leaves at seedling stage. The established method potentially can be used to monitor PMI protein in rice grains.
文摘目的分析张家港市2018年检出的65株沙门菌的血清型特征及分布特点,探讨该地区沙门菌优势菌群的变迁。方法收集张家港市2018年来自体检、食物中毒、食源性疾病主动监测等人源性沙门菌65株,运用脉冲场凝胶电泳(Pulsed Field Gel Electrophoresis,PFGE)技术对其中优势血清群进行分子分型,利用PulseNet China网络获得相关性指纹图谱并进行聚类分析。结果65株沙门菌分离株分布6个血清群和9个血清型,以D群为主(46.97%)、B群次之(21.21%)。31株D群沙门菌分为6个基因型(相似值100.00%为同一PFGE型别),条带相似度为71.63%~100.00%,2个克隆系(相似度为0.85~1.00的为同一PFGE的克隆系),2个克隆系T1与T2相似度为71.63%,克隆系相似度较高。14株B群沙门菌PFGE指纹图谱可分为12个克隆系13种基因型,条带相似度为10.21%~100.00%。结论张家港市2018年人源沙门菌血清型呈多样化分布,优势血清群D群PFGE带型存在较近亲缘关系,B群沙门菌PFGE带型较多,基因多态性较高。
