Table 1 Influence of type of donor DNA (dDNA) (Lg-ss-dDNAs, Ld-ss-dDNAs, ds-dDNA) length of dDNA, concentration of dDNA and size of deletion on mutagenesis efficiency at four chromosomal loci
Target gene-pCRISPR-gDNA | Type of dDNA | dDNA ID | dDNA length (nt) | Mutation (nt) | dDNA quantity (µg) | Efficiency (%)-positive/total |
|---|---|---|---|---|---|---|
ompF 5′-pCRISPR-ompF_5′G | ss-Lg | ompF_5′G-70-∆30 | 70 | ∆30 | 1 | 86% (19/22) |
70 | ∆30 | 10 | 95% (40/42) | |||
ompF_5′G-70-∆100 | 70 | ∆100 | 10 | 5% (2/36) | ||
ompF_5′G-70-∆500 | 70 | ∆500 | 10 | 0% (0/30) | ||
ompF_5′G-120-∆30 | 120 | ∆30 | 1 | 64 ± 34% (40/62) | ||
120 | ∆30 | 10 | 93% (76/82) | |||
ompF_5′G-120-∆100 | 120 | ∆100 | 10 | 64 ± 31% (20/31) | ||
ompF_5′G-120-∆500 | 120 | ∆500 | 10 | 47 ± 19% (14/30) | ||
ompF_5′G-120-∆1089 | 120 | ∆1180 | 10 | 0% (0/20) | ||
ss-Ld | ompF_5′G-70-∆30 R | 70 | ∆30 nt | 10 | 20% (4/20) | |
ompF_5′G-120-∆30 R | 120 | ∆30 | 10 | 100% (10/10) | ||
ompF_5′G-120-∆1089 R | 120 | ∆1180 | 10 | 0% (0/> 100) | ||
ds | ompF_5′G-120-∆30 ds | 120 | ∆30 | 10 | 77% (20/26) | |
ompF_5′G-120-∆500 ds | 120 | ∆500 | 10 | 50 ± 14% (15/30) | ||
ompF_5′G-120-∆1089 ds | 120 | ∆1180 | 10 | 26 ± 17% (6/23) | ||
ompF 3′-pCRISPR-ompF_3′I | ss-Lg | ompF_3′I-70-∆30 | 70 | ∆30 | 10 | 79% (19/24) |
ompF_3′I-120-∆30 | 120 | ∆30 | 10 | 83% (25/30) | ||
fecA-pCRISPR-fecA_B | ss-Lg | fecA_B-70-∆30 | 70 | ∆30 | 10 | 43 ± 4% (13/30) |
fecA_B-120-∆30 | 120 | ∆30 | 10 | 77% (23/30) | ||
fecA_B-120-∆100 | 120 | ∆100 | 10 | 20% (6/30) | ||
fecA_B-120-∆500 | 120 | ∆500 | 10 | 13% (4/30) | ||
fecA_B-120-∆2325 | 120 | ∆2325 | 10 | 0% (0/30) | ||
ds | fecA_B-120-∆2325 ds | 120 | ∆2325 | 10 | 14% (5/35) | |
lpp-pCRISPR-lpp_B | ss-Lg | lpp_B-70-∆30 R | 70 | ∆30 | 10 | 20% (4/20) |
lpp_B-120-∆30 R | 120 | ∆30 | 10 | 55 ± 7% (11/20) | ||
lpp_B-120-∆237 R | 120 | ∆237 | 10 | 0% (0/16) | ||
ss-Ld | lpp_B-70-∆30 | 70 | ∆30 | 10 | 90% (27/30) | |
lpp_B-120-∆30 | 120 | ∆30 | 10 | 73% (19/26) | ||
lpp_B-120-∆237 | 120 | ∆237 | 10 | 0% (0/30) | ||
ds | lpp_B-120-∆30 ds | 120 | ∆30 | 10 | 82% (19/23) | |
lpp_B-120-∆237 ds | 120 | ∆237 | 10 | 72% (18/25) |