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    • 3. 发明授权
    • Inviable phages, their production and DNA thereof
    • 不可侵犯的噬菌体,其生产和DNA
    • US5559018A
    • 1996-09-24
    • US946033
    • 1992-09-15
    • Thomas L. MattsonRichard Epstein
    • Thomas L. MattsonRichard Epstein
    • C12N15/70C12N15/00C12N15/63
    • C12N15/70
    • Inviable T4 phage-like particles capable of directing the expression of large non-T4 DNA fragments from T4 expression control sequences are produced. Thus, E. coli harboring pBR322 derivatives containing cloned T4 gene 23 DNA sequences were infected with T4 phage carrying a deletion of the denB gene. Homology-dependent recombination results in the production of inviable phage-like particles containing DNA molecules composed of multiple, tandemly repeated copies of entire plasmid molecules covalently linked to single copies of normal phage genes. The yield of these inviable particles, intially low, was increased by means of a reiterated infection process that involves the use of a cloned T4 origin of replication. When T4 gene 32 expression control sequences linked in proper orientation to a DNA sequence coding for the non-T4 protein .beta.-galactosidase were also cloned in one such pBR322 derivative (pVH773), inviable phage particles capable of directing the synthesis of enzymatically active .beta.-galactosidase were produced. The present process is applicable to other T-even bacterio-phages.
    • 产生能够引导来自T4表达调控序列的大的非T4 DNA片段表达的不可维生的T4噬菌体样颗粒。 因此,含有克隆的T4基因23个DNA序列的pBR322衍生物的大肠杆菌用携带denB基因缺失的T4噬菌体感染。 同源性依赖性重组导致产生含有DNA分子的易发生的噬菌体样颗粒,其由与正常噬菌体基因的单拷贝共价连接的整个质粒分子的多个串联重复拷贝组成。 通过涉及使用克隆的T4复制起点的重复的感染过程,这些不可侵犯的颗粒的产量是非常低的。 当将T4基因32表达控制序列与编码非T4蛋白质β-半乳糖苷酶的DNA序列连接的表达调控序列也克隆在一个这样的pBR322衍生物(pVH773)中时,能够引导酶活性β- 产生半乳糖苷酶。 本方法适用于其他T-even细菌噬菌体。