TABLE 1.
Strains, plasmids, and oligonucleotides used in this study
| Strain, plasmid, or oligonucleotide | Descriptiona | Reference |
|---|---|---|
| E. coli strains | ||
| BW25113 | rrnBT14 ΔlacZWJ16hsdR514 ΔaraBADAH33 ΔrhaBADLD78 | 7 |
| JCL16 | BW25113 F′ [traD36proAB+lacIqZΔM15] | 2 |
| JCL260 | Same as JCL16 but ΔadhE Δfnr-ldhA ΔfrdBC ΔpflB Δpta | 3 |
| SA296 | Same as JCL260 but ΔilvC | This work |
| KS145 | Same as JCL16 but ΔilvI ΔilvB | 4 |
| Plasmids | ||
| pSA55 | ColE1 ori; Ampr; PLlacO1::kivd-ADH2 | 3 |
| pSA69 | p15A ori; Kanr; PLlacO1::alsS-ilvC-ilvD | 3 |
| pCS27 | p15A ori; Kanr; PLlacO1::MCS1 | 16 |
| pZL8 | p15A ori; Kanr; PLlacO1::alsS | This work |
| pSA159 | Derivative of pPETDuet-1 with alsS | This work |
| pSA166 | Derivative of pPETDuet-1 with alsS(Q487A) | This work |
| pSA187 | Derivative of pPETDuet-1 with alsS(Q487I) | This work |
| pSA188 | Derivative of pPETDuet-1 with alsS(Q487S) | This work |
| pSA205 | Derivative of pPETDuet-1 with alsS(Q487G) | This work |
| pSA206 | Derivative of pPETDuet-1 with alsS(Q487L) | This work |
| Oligonucleotides | ||
| A124 | ACGCAGTCGACCTAGAGAGCTTTCGTTTTCATGAGT | 3 |
| A297 | CGGGATCCGTTGACAAAAGCAACAAAAGAACAAA | This work |
| A298 | ACGCAGTCGACCTAGAGAGCTTTCGTTTTCATGAGT | This work |
| A300 | AATAAGACGTCTAAGAAACCATTATTATCATG | This work |
| A305 | GAATGCAACCATGTCATATGTGCTG | This work |
| A306 | ATGGAACGACAGCACATATGACATGGTTGCATTCAACCAATTGAAAAAATATAACCGTAC | This work |
| A307 | ATGGAACGACAGCACATATGACATGGTTGCATTCGCCCAATTGAAAAAATATAACCGTAC | This work |
a
For oligonucleotides, the sequences are shown (5′→3′).