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。
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.
出处
《潍坊医学院学报》
1994年第2期81-85,共5页
Acta Academiae Medicinae Weifang
