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    • 4. 再颁专利
    • Molecular genetic construction of vaccine strains of pasteurellaceae
    • 巴氏杆菌科疫苗株的分子遗传构建
    • USRE38028E1
    • 2003-03-11
    • US09716319
    • 2000-11-21
    • Robert E. BriggsFred M. Tatum
    • Robert E. BriggsFred M. Tatum
    • C12N100
    • C07K14/285A61K39/00C12N9/1007C12N9/22C12N15/74
    • Tools for genetically engineering Pasteurellaceae are provided. Replication-conditional plasmids which are useful for the Pasteurellaceae have been isolated and characterized. The plasmids can be utilized for delivery of DNA segments into the Pasteurellaceae in situations where control of extra-chromosomal replication desired, such as in achieving allelic exchange or site-directed mutagenesis. A restriction endonuclease, HsoI, was isolated from a bovine lung isolate of Haemophilus somnus. The enzyme was found to be a true isoschizomer of HinPI, a commercially available enzyme originally isolated from Haemophilus influenzae PI. Commercially available HhaI methyl transferase was found to protect against cleavage by both enzymes. Methylation of foreign plasmid DNA was found to enhance transformation of Haemophilus somnus in excess of four orders of magnitude.
    • 提供了基因工程巴斯德氏菌科的工具。 已经分离和表征了可用于巴氏杆菌科的复制条件质粒。 在需要控制染色体外复制的情况下,例如在实现等位基因交换或定点诱变的情况下,可以将质粒用于将DNA片段递送至巴氏杆菌科。将限制性内切核酸酶HsoI从牛肺分离株 嗜血嗜血虫 该酶被发现是HinPI的真正的同位异构体,HinPI是最初从流感嗜血杆菌PI中分离的市售酶。 发现市售的HhaI甲基转移酶可以防止两种酶切割。 发现外源质粒DNA的甲基化可增强嗜睡嗜血杆菌的转化,超过四个数量级。