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椰毒假单胞菌部分16s rRNA序列测定法

A SEQUENCING METHOD OF PSEUCOMONAS COCOVENENANS PARTIAL 16s rRNA

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【作者】 曲春枫李在连赵乃昕汪恩涛陈文新

【Author】 Qu Chunfeng; Li Zailian; Zhao Naixin; et al (Department of Immunology, 261042)

【机构】 北京农业大学生物学院

【摘要】 详细介绍了椰毒假单胞菌部分16srRNA序列测定法。常规提取细菌DNA,采用聚合酶链反应(PCR)扩增目的基因,琼脂糖凝胶电泳后回收目的基因。纯化后克隆到M13mp19噬菌体上,转染大肠杆菌JM109菌株。挑选无色噬斑扩增并提取其双链DNA,酶切分析为阳性克隆,提取其单链DNA在ABI公司370A型自动测序仪上测序。

【Abstract】 A sequencing method of Pseudomonas Cocovenenans partial 16s rRNA was introduced in details in our paper. The target gene was amplified by polymerase chain reaction and recovered by agrose gel electrophoresis. After purified, the target gene was cloned to vector bacteriophage M13 mp19 to transfect E. Coli JM109.There were 15 colourless peaques in all. Selected one of the plaques and propagated it in LB broth. The recombinant M14mp19 double strand DNA was extracted to be anaysed by restrictive enzymes EcoR I and Hind. When the seleted clone was defined to be positive, the single stranded DNA was extracted and sequenced in the model 370 A automated DNA sequencer.

  • 【文献出处】 潍坊医学院学报 ,JOURNAL OF WEIFANG MEDICAL COLLEGE , 编辑部邮箱 ,1994年02期
  • 【分类号】R329.26
  • 【被引频次】1
  • 【下载频次】74
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