TABLE 6.
N. tabacum transformation with different tra(N)-mobilizable vectorsa
| Donor strain(s) | Total no. of tumors/total no. of leaf strips (avg) |
|---|---|
| Direct coinoculation | |
| DH5α(pKM101, pKP10) + EHA101 | 119/49 (2.4) |
| DH5α(pKM101, pKP50) + EHA101 | 7/64 (0.13) |
| DH5α(pKM101, pKP80) + EHA101 | 82/40 (2.05) |
| DH5α(pKM101, pKP80ΔN) + EHA101 | 22/55 (0.4) |
| DH5α(pKM101, pKP81) + EHA101 | 336/52 (0.7) |
| Strains filter mated prior to plant inoculationb | |
| DH5α(pKM101, pKP10) + EHA101 | 228/35 (6.5) |
| DH5α(pKM101, pKP80ΔN) + EHA101 | 58/42 (1.3) |
| DH5α(pKM101, pKP81) + EHA101 | 67/38 (1.7) |
| A. tumefaciens control donor strains | |
| EHA101(pKP10) | 1,440/21 (68.5) |
| EHA101(pKP50) | 1,172/15 (78.1) |
| EHA101(pKP80) | 880/12 (73.3) |
a
All vectors were mobilized from the E. coli donor strains to E. coli or A. tumefaciens recipients with similar frequencies in filter matings (data not shown). E. coli DH5α(pKM101) strains harboring each of the vectors on the left were used to transform tobacco strips in the presence of helper A. tumefaciens strain EHA101.
b
Leaf strip infection followed filter matings of the DH5α(pKM101) donors harboring the relevant vectors and EHA101 on LB agar plates for 5 h (see Materials and Methods).