Table 1.
Methods for the integration of large DNA module into E. coli chromosome
| Method | Technique feature | Integration efficiency (%) | Integration sites | Maximum integration size (kb) | Rounds of integration | Marker left or markerless |
Reference |
|---|---|---|---|---|---|---|---|
| I-SceI endonuclease | Combined λ-Red and I-SceI Cleavage | 19.2–100 | Desired locus | 7 | One | Markerless | [5] |
| λ-Red recombination | λ-Red assisted | 0–50 | Desired locus | 7.3 | One | Markerless | [7] |
| λ-Red recombination | Combined λ-Red and CRISPR/Cas9 | 10 | Desired locus | 7 | One | Markerless |
[33] [9] |
| 60 | 15.4 | Five | Markerless | ||||
| λ-Red recombination | λ-Red assisted | Not provide | fliK | 15 | Four | Markerless | |
| Not provide | fliK | 50 | Seven | Markerless | |||
| CRISPR/Cas9 | Combined λ-Red and CRISPR/Cas9 | 60 | Desired locus | 7 | One | Markerless | [12] |
| CRISPR/Cas9 | Combined λ-Red and CRISPR/Cas9 | 50 | Desired locus | 10 | One | Markerless | [13] |
| RecA homologous recombination | Combined λ-Red, RecA and CRISPR/Cas9 | 100 | IS5 | 12 | One | Markerless | This study |