TABLE 1.
Strains, plasmids, and oligonucleotide primers used in this study
| Strain, plasmid, or primer | Relevant phenotype and description or sequence (5′-3′)a | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli strains | ||
| VE7108 | Kanamycin resistance marker; contains the wild-type repA plasmid gene (not thermosensitive); host for pG+host9 DNA manipulation | 12 |
| VE6838 | Kanamycin resistance marker; VE7108 carrying pG+host9 | 12 |
| Lactobacillus plantarum strains | ||
| WT | NCIMB 8826 WT strain | NCIMB |
| ΔldhL1 | NCIMB 8826 ΔldhL1 strain | This study |
| ldhL1::amyA | NCIMB 8826 strain with ldhL1 gene replaced with amyA-secreting expression cassette | This study |
| Plasmids | ||
| pCUSαA | Vector for secreting α-amylase; erythromycin resistance marker | 16 |
| pG+host9 | Thermosensitive replicon; erythromycin resistance marker | 10 |
| pGh9-ΔldhL1 | Vector for deletion of ldhL1 gene of NCIMB 8826; erythromycin resistance marker | This study |
| pGh9-ldhL1::amyA | Vector for replacement of ldhL1 gene of NCIMB 8826 with amyA-secreting expression cassette; erythromycin resistance marker | This study |
| Oligonucleotide primers | ||
| ldhL1-up_F | CCGCTCGAGAATCCGGTGATTAGTTGCCG | |
| ldhL1-up_R | ACGCGTCGACGAGCTCAGATCTAATAAGTCATCCTCTCGTAGTGAAAA | |
| ldhL1-down_F | CCCGATATCGTCGACTCATTTCATACGATTAAATGTATGATGAACG | |
| ldhL1-down_R | GGACTAGTAACAGCGTTTTGGTCTAAACTTACTTT | |
| ldhL1-up_seq | AAATCTTCTCACCGTCTTGAAAAG | |
| ldhL1-down_seq | TGGTTAAATCTTTTTTAGTATTGATTGTAATACC |
a
For primers, restriction enzyme sites are underlined.