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    • 6. 发明授权
    • Bacteriophage resistant recombinant bacteria
    • 耐噬菌细菌重组菌
    • US5538864A
    • 1996-07-23
    • US373877
    • 1995-01-17
    • Colin J. HillTodd R. Klaenhammer
    • Colin J. HillTodd R. Klaenhammer
    • A23C9/123C12N1/04C12N1/21C12N15/11C12N15/34C12N15/65C12N15/74C12N1/20A23C9/00C12P7/56
    • C12N15/65A23C9/1236C12N1/04C12N15/11C12N15/746
    • Recombinant bacteria containing phage-encoded resistance ("Per") and methods of making and using the same are disclosed. Such bacteria are made by (a) conducting a fermentation of a substrate in a medium containing a defined bacterial culture until bacteriophage are detected in the medium, the bacteriophage being specific to at least one bacteria in the medium; (b) isolating the bacteriophage; (c) digesting DNA of the bacteriophage to produce a library of DNA fragments; (d) transforming the bacteria susceptible to said bacteriophage with the library of DNA fragments to provide transformed bacteria; (e) selecting from among the transformed bacteria, a bacteriophage-resistant transformed bacteria; (f) adding bacteriophage resistant transformed bacteria to the medium; and (g) recommencing step (a). Also disclosed are bacterial cells which contain a first bacteriophage defense mechanism (Per), wherein Per comprises a bacteriophage origin of replication (ori) operatively associated with a DNA sequence incapable of producing live bacteriophage. The bacterial cell is capable of being infected by a bacteriophage, the DNA of which, once injected into the bacterial cell, competes with Per for binding to DNA polymerase.
    • 公开了含有噬菌体编码抗性(“Per”)的重组细菌及其制备和使用方法。 这样的细菌通过以下方法制备:(a)在含有限定的细菌培养基的培养基中进行底物的发酵,直到在培养基中检测到噬菌体,噬菌体对培养基中的至少一种细菌是特异性的; (b)分离噬菌体; (c)消化噬菌体的DNA以产生DNA片段文库; (d)用DNA片段的文库转化对所述噬菌体易感的细菌,以提供转化的细菌; (e)从转化细菌中选择抗噬菌体转化细菌; (f)向培养基中加入抗噬菌体转化细菌; 和(g)重新开始步骤(a)。 还公开了含有第一噬菌体防御机制(Per)的细菌细胞,其中Per包含与不能产生活噬菌体的DNA序列可操作地相关的噬菌体复制起点(ori)。 细菌细胞能够被噬菌体感染,其噬菌体一旦被注入到细菌细胞中就与Per结合就与DNA聚合酶结合。