Table 3 Recovery of functional GroEL-variants and Adk-variants from mock libraries with increasing excess of non-functional variants using the established selection system in comparison with a conventional bleach-out system
From: A genetic replacement system for selection-based engineering of essential proteins
Strain | Library (functional: non-functional) | Number of colonies after transformationa | Number of genotyped variants | Number of detected false positive variants |
|---|---|---|---|---|
W3110 groE::kan [pCOMP-groE, pP ara I –Sce I] | Only functional | ~ 27 000 | 20 | 0 |
1:102 | 231 | 20 | 0 | |
1:103 | 33 | 20 | 0 | |
1:104 | 4 | 4 | 0 | |
1:105 | 1a | 1 | 0 | |
Only non-functional | 0 | 0 | 0 | |
SBΔrecA | Only functional | ~ 25 000 | 20 | 0 |
1:102 | 253 | 20 | 0 | |
1:103 | 25 | 20 | 1 | |
1:104 | 7 | 7 | 5 | |
1:105 | 6 | 6 | 5 | |
Only non-functional | 7 | 7 | 7 | |
W3110 adk::kan [pCOMP-adk, pP ara I-Sce I] | Only functional | ~ 46 000 | 20 | 0 |
1:102 | 459 | 20 | 0 | |
1:103 | 36 | 20 | 0 | |
1:104 | 3 | 3 | 0 | |
1:105 | 1 | 1 | 0 | |
Only non-functional | 0 | 0 | 0 |