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Fig. 1 | Microbial Cell Factories

Fig. 1

From: Copy number variability of expression plasmids determined by cell sorting and Droplet Digital PCR

Fig. 1

Design of plasmid vectors and experimental strategy. a Genetic map of p2X4-AEB plasmid vector series. Seven different replication systems (X) obtained from the SEVA repository were cloned into a typical expression vector, pSEVA2X4-StyA-EGFP StyB (p2X4-AEB). Listed in the table is the official SEVA module number (2–8), name and structure of the replication system (orange—replication initiation proteins, blue—origin of replication), and size. The map illustrates the genetic structure of the plasmid backbone including the lacI q repressor, the tryp-lac hybrid promoter (P trc ), a styrene monooxygenase (styA) in frame fused to an EGFP reporter, an FAD cofactor reductase (styB), a kanamycin resistance gene (kan R) and the origin of transfer (oriT). bp—base pairs. b Cell sorting and digital PCR for copy number determination. 1—bacterial cultivation, 2—population analysis by flow cytometry, 3—sorting 1000 cells/well, 4—DNA extraction and droplet formation, 5—Droplet Digital PCR reaction, 6—counting positive and negative droplets. Adapted with permission from [7]. Copyright (2014) American Chemical Society

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