TABLE 3.
Results of gene disruption recombineering applied to antitermination and ribosome-associated protein genes
| Gene | Recombination efficiencya | Gene configurationb | Gene essentiality |
|---|---|---|---|
| nusA | 1.8 × 101 | nusA<>cat/nusA+ | Essential |
| nusBc | 0.9 × 101 | nusB<>cat/nusB+ | NAe |
| 6.5 × 103 | nusB<>cat | Growth impaired | |
| nusD/rho | 3.7 × 102 | nusD<>kan/nusD+ | Essential |
| rhoL-nusD/rho | 2.7 × 102 | nusD<>kan/nusD+ | Essential |
| rhoL-nusD/rhod | 4.2 × 102 | nusD<>kanSD/nusD+ | Essential |
| nusE(rpsJ) | 0.7 × 101 | nusE<>kan/nusE+ | Essential |
| nusG | 1.2 × 101 | nusG<>cat/nusG+ | Essential |
| yfiA | 7.3 × 104 | yfiA<>kan | Nonessential |
| yhbH | 6.6 × 104 | yhbH<>cat | Nonessential |
| rmf | 3.2 × 104 | rmf<>amp | Nonessential |
| rpsV | 8.5 × 104 | rpsV<>kan | Nonessential |
a
Number of antibiotic-resistant DY330 recombinants per 5 × 108 viable cells.
b
Determined by PCR analysis of the chromosomal region with the replaced gene, as shown in Fig. 3. Partial diploids are shown as gene<>antibiotic-resistance cassette/gene+.
c
Two gene knockout patterns were observed for nusB knockouts. Cells with a diploid gene configuration appeared with a low frequency on the first day. After 2 days, they were followed by numerous nusB<>cat recombinants.
d
rhoL-rho was replaced with kan containing its own Shine-Dalgarno sequence, kanSD.
e
NA, not applicable.