FILAMENTOUS PHAGE DNA CLONING VECTORS - A NON-INFECTIVE MUTANT WITH A NON-POLAR DELETION IN GENE-III
FILAMENTOUS PHAGE DNA CLONING VECTORS - A NON-INFECTIVE MUTANT WITH A NON-POLAR DELETION IN GENE-III
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DOI：
10.1016/0042-6822(81)90442-6
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发表时间：
1981-01-01
期刊：
影响因子：
3.7
通讯作者：
SMITH, GP
中科院分区：
文献类型：
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作者：
NELSON, FK;FRIEDMAN, SM;SMITH, GP
以四环素抗性转导噬菌体fd-tet为基础，通过删除噬菌体基因III的一个50碱基对（bp）片段，构建了丝状噬菌体衍生物fKN16。与基因III蛋白在感染中的重要性相一致，fKN16噬菌体的传染性小于fd-tet噬菌体的10-8。大多数III基因琥珀色突变体在下游噬菌体基因VI和I上是极性的，与之相反，fKN16应该是一个非极性突变体，因为其507 bp的缺失跨越了整数个编码三联体。两种可能依赖于基因VI和I功能的噬菌体特征，即噬菌体产生和包装成单位长度病毒粒子的水平，在fKN16中似乎是正常的。高噬菌体产量加上非常低的传染性[大肠杆菌]使fKN16适合作为需要高水平生物遏制的DNA克隆实验的载体。利用噬菌体.lambda的HindIII片段，详细研究了fKN16作为载体的特性。DNA作为外源DNA模型。fKN16可能有助于研究基因III蛋白在丝状噬菌体生命周期中的作用。
A filamentous phage derivative, fKN16, was constructed from the tetracycline-resistance transducing phage fd-tet by deleting a 50 base-pair (bp) segment of phage gene III. In accord with the importance of the gene III protein in infection, the infectivity of fKN16 phage is < 10-8 that of fd-tet phage. In contrast to most gene III amber mutants, which are polar on the downstream phage genes VI and I, fKN16 should be a nonpolar mutant since its 507 bp deletion spans an integral number of coding triplets. Two phage traits that may depend on gene VI and I function, the level of phage production and packaging into unit-length virions, appear to be normal in fKN16. High phage production coupled with very low infectivity [Escherichia coli] make fKN16 suitable as a vector for DNA cloning experiments requiring a high level of biological containment. The characteristics of fKN16 as a vector were investigated in detail, using HindIII fragments of phage .lambda. DNA as model foreign DNA. fKN16 may be useful in studying the role of the gene III protein in the filamentous phage life cycle.