Filamentous phage infection-mediated gene expression: construction and propagation of the gIII deletion mutant helper phage R408d3
Filamentous phage infection-mediated gene expression: construction and propagation of the gIII deletion mutant helper phage R408d3
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DOI：
10.1016/s0378-1119(97)00298-9
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发表时间：
1997-10-01
期刊：
影响因子：
3.5
通讯作者：
Model, P
中科院分区：
文献类型：
--
作者：
Rakonjac, J;Jovanovic, G;Model, P
我们描述了使用的噬菌体休克蛋白（PSP）启动子的转录融合。这些融合蛋白仅在细胞被丝状噬菌体感染时表达。在一个应用中，将psp启动子融合到丝状噬菌体基因III（gIII）的蛋白质编码部分。需要蛋白III（pIII）来补充含有gIII缺失的突变体fl噬菌体，但其合成也使细胞对感染具有抗性。通过仅在已经被噬菌体感染的细胞中诱导psp-gIII产生pIII，可以从其中gIII已经完全缺失的噬菌体中获得噬斑。gIII从两个辅助噬菌体：R408和VCSM 13中缺失，然后在含有psp-gIII融合体的细胞上繁殖。测试这两种质粒用于噬菌体展示方法，该方法需要产生非感染性的含噬菌粒的病毒体样颗粒。这两种辅助病毒都有效，但R408 d3比VCSM 13 d3更上级，因为它产生的背景感染性颗粒少1800倍。(C)1997年Elsevier Science B.V.
We describe the use of transcriptional fusions to the phage shock protein (psp) promoter. These fusions are expressed only when cells are infected by filamentous phage. In an application, the psp promoter was fused to the protein coding part of filamentous phage gene III (gIII). Protein III (pIII) is needed to complement mutant fl phage containing a deletion of gIII but its synthesis also renders cells resistant to infection. By inducing pIII production from psp-gIII only in the cells that are already infected with phage, it was possible to obtain plaques from phage in which gill had been completely deleted.gIII was deleted from two helper phages: R408 and VCSM13, which were then propagated on cells containing the psp-gIII fusion. These two phages were tested for use in a phage display method that requires generation of noninfectious, phagemid-containing virion-like particles. Both helpers worked, but R408d3 was superior to VCSM13d3, because it generated about 1800-times fewer background infectious particles. (C) 1997 Elsevier Science B.V.