Table 2 Transformation efficiencies and frequency of false positive variants for the Adk-specific selection systems
From: A genetic replacement system for selection-based engineering of essential proteins
Strain | I-Sce I induction during competent cell preparation | Number of transformants (frequency CFU per transformant) | ||
|---|---|---|---|---|
pSEVA132-adk | pSEVA132-adkstop | pSEVA132 | ||
W3110 adk::kan [pKOCOMP- adk , pI- Sce I] | No | 20 200 ± 2900 | 66 ± 21 (~3x10-3) | 28 ± 13 (~1x10-3) |
Yes | 21 000 ± 6000 | 0 (<2x10-4) | 0 (<2 x10-4) | |
W3110 adk ::kan [pCOMP- adk, pP ara I- Sce I] | No | 32 100 ± 3700 | 61 ± 17 (2x10-3) | 35 ± 11 (1x10-3) |
Yes | 28 000 ± 2200 | 0 (<2x10-4) | 0 (<2x10-4) | |