The role of the absorption complex in the termination of filamentous phage assembly

Gailus, Valérie; Ramsperger, Uwe; Johner, Christian; Krämer, H.; Rasched, Ihab

doi:10.1016/0923-2508(94)90042-6

Cited by 7 publications

(1 citation statement)

References 32 publications

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“…To enable the host cells to control the infectivity of packaged phages, we transfer an essential gene for phage infectivity from the M13 phage to the host cells. The essential gene applied in this study is gene III encoding the attachment protein pIII which mediates adsorption of the phage to its primary receptor, the tip of E. coli F-pilus 30 . We then design an intracellular biological circuit to control the infectivity of packaged phages by controlling the expression level of gene III based on a specific intracellular signal that is related to the performance of the biomolecules to be screened, such as the concentration of an end product or an intermediate metabolite of a metabolic pathway.…”

Section: Resultsmentioning

confidence: 99%

Engineering ‘cell robots’ for parallel and highly sensitive screening of biomolecules under in vivo conditions

Song

Zeng

2017

Sci Rep

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Cells are capable of rapid replication and performing tasks adaptively and ultra-sensitively and can be considered as cheap “biological-robots”. Here we propose to engineer cells for screening biomolecules in parallel and with high sensitivity. Specifically, we place the biomolecule variants (library) on the bacterial phage M13. We then design cells to screen the library based on cell-phage interactions mediated by a specific intracellular signal change caused by the biomolecule of interest. For proof of concept, we used intracellular lysine concentration in E. coli as a signal to successfully screen variants of functional aspartate kinase III (AK-III) under in vivo conditions, a key enzyme in L-lysine biosynthesis which is strictly inhibited by L-lysine. Comparative studies with flow cytometry method failed to distinguish the wild-type from lysine resistance variants of AK-III, confirming a higher sensitivity of the method. It opens up a new and effective way of in vivo high-throughput screening for functional molecules and can be easily implemented at low costs.

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Section: Resultsmentioning

confidence: 99%