Table 2 Oligonucleotides used in this study
Oligonucleotide name | Sequence (5′ → 3′) | Purpose |
|---|---|---|
cg1497_upstrm_fw_pK19 | GACTCTAGAGGATCCCCTTAACGCGCCGGGCTC | pK19mobsacB-Δcg1497 |
cg1497_upstrm_rv | GGGTAGGTGATTTGAATTTGTGCTTTCGGAACTGGACATAATCAGATAC | pK19mobsacB-Δcg1497 |
cg1497_dwnstrm_fw | ACAAATTCAAATCACCTACCCGGAATGGAGAATCTGGTAGAGATCGG | pK19mobsacB-Δcg1497 |
cg1497_dwnstrm_rv_pK19 | CGAGCTCGGTACCCGAACTCTGGATGAGATAGCTGAGGTT | pK19mobsacB-Δcg1497 |
Dcg1497_fw_v3 | CCACTGCCACGGAGCC | Verification of cg1497 deletion by PCR |
Dcg1497_rv_v3 | AACGAAGTGCCACTTCTTCCAC | Verification of cg1497 deletion by PCR |
nagD_upstrm_fw_pK19 | GACTCTAGAGGATCCCCTTCCCCGCAATGAGCCG | pK19mobsacB-ΔhdpA |
nagD_upstrm_rv | GGGTAGGTGATTTGAATTTGTTGAAATGTTCACTGTCATAACACCATTGT | pK19mobsacB-ΔhdpA |
nagD_dwnstrm_fw | ACAAATTCAAATCACCTACCCTTTCACGTACCAGATGAGCAGC | pK19mobsacB-ΔhdpA |
nagD_dwnstrm_rv_pK19 | CGAGCTCGGTACCCGGAACCTTCGGCTTGGATCTG | pK19mobsacB-ΔhdpA |
DnagD_fw | GATGAACACGACCGTTGCC | Verification of hdpA deletion by PCR |
DnagD_rv | GGGTGGTCTTTGAGGAGTTCTTC | Verification of hdpA deletion by PCR |
ldhfow | TGATGGCACCAGTTGCGATGT | Verification of ldh deletion by PCR |
ldhrev | CCATGATGCAGGATGGAGTA | Verification of ldh deletion by PCR |
mgsA_fw_x3 | GACTCTAGAGGATCCCC GAAAGGAGGCCCTTCAGATGGAACTGACGACTCGCACT | pEKEx3-mgsA-gldA, pEKEx3-mgsA-yqhD-gldA, pEKEx3-mgsA-yqhD-fucO-gldA |
mgsA_rv_gld_DS | TATCTCATAAAGTTACTTCAGACGGTCCGCGA | pEKEx3-mgsA-gldA |
gldA_fw_mgs_DS | GGACCGTCTGAAGTAACTTTATGAGATA GAAAGGAGGCCCTTCAGATGGACCGCATTATTCAATCACCG | pEKEx3-mgsA-gldA |
gldA_rv_x3 | CGAGCTCGGTACCCTTATTCCCACTCTTGCAGGAAAC | pEKEx3-mgsA-gldA, pEKEx3-mgsA-yqhD-gldA, pEKEx3-mgsA-yqhD-fucO-gldA |
mgsA_rv | TTACTTCAGACGGTCCGCGA | pEKEx3-mgsA-yqhD-gldA |
yqhD_fw_mgs | GGACCGTCTGAAGTAA GAAAGGAGGCCCTTCAGATGAACAACTTTAATCTGCACACCCC | pEKEx3-mgsA-yqhD-gldA |
yqhD_rv | TTAGCGGGCGGCTTCGTATATA | pEKEx3-mgsA-yqhD-gldA |
gldA_fw_yqh | GCCGCCCGCTAA GAAAGGAGGCCCTTCAGATGGACCGCATTATTCAATCACCG | pEKEx3-mgsA-yqhD-gldA |
mgsA_rv_yqh_DS | TATCTCATAAAGTTACTTCAGACGGTCCGCGA | pEKEx3-mgsA-yqhD-fucO-gldA |
yqhD_fw_mgs_DS | GGACCGTCTGAAGTAACTTTATGAGATA GAAAGGAGGCCCTTCAGATGAACAACTTTAATCTGCACACCCC | pEKEx3-mgsA-yqhD-fucO-gldA |
yqhD_rv_gld_DS | GAAATGAATAGCTTAGCGGGCGGCTTCGTATATA | pEKEx3-mgsA-yqhD-fucO-gldA |
fucO_fw_yqh_DS | GCCGCCCGCTAAGCTATTCATTTC GAAAGGAGGCCCTTCAGATGATGGCTAACAGAATGATTCTGAACG | pEKEx3-mgsA-yqhD-fucO-gldA |
fucO_rv_gld_DS | AAGGCAAGAATCTTACCAGGCGGTATGGTAAAGCT | pEKEx3-mgsA-yqhD-fucO-gldA |
gldA_fw_fuc_DS | CATACCGCCTGGTAAGATTCTTGCCTT GAAAGGAGGCCCTTCAGATGGACCGCATTATTCAATCACCG | pEKEx3-mgsA-yqhD-fucO-gldA |
ppdABC_ko_fw_x1 | CTGCAGGTCGACTCTAGAG GAAAGGAGGCCCTTCAGATGAGATCGAAAAGATTTGAAGCACTGG | pVWEx1-ppdABC |
ppdABC_ko_rv_x1 | CGGTACCCGGGGATCTTAATCGTCGCCTTTGAGTTTTTTACG | pVWEx1-ppdABC |
gldA_Seq | GAACTGTGCTACAACACCCTG | Sequencing primer for gldA |
yqhD_Seq | GTATTTGCCGTGCTCGATC | Sequencing primer for yqhD |
fucO_Seq | GACCAATAAACCCAGTGTAC | Sequencing primer for fucO |
ppdABC_Seq1 | CGAACAGGAAACCACCGTTG | Sequencing primer for ppdABC |
ppdABC_Seq2 | ACGACCAGACCTTCACCCAC | Sequencing primer for ppdABC |
ppdABC_Seq3 | TACCTGCATACCTCCGCGAT | Sequencing primer for ppdABC |
ppdABC_Seq4 | AATCCTCCGACGTGGCCTTC | Sequencing primer for ppdABC |
ppdABC_Seq5 | CGAACAAGCACCCGGAATGG | Sequencing primer for ppdABC |
pVWEx1_fw | CATCATAACGGTTCTGGC | Verification of correct pEKEx3/pVWEx1 derivatives by PCR/sequencing |
pVWEx1_rv | ATCTTCTCTCATCCGCCA | Verification of correct pEKEx3/pVWEx1 derivatives by PCR/sequencing |
M13_fw | CGCCAGGGTTTTCCCAGTCACGAC | Verification of correct pK19mobsacB derivatives by PCR/sequencing |
M13_rv | AGCGGATAACAATTTCACACAGGA | Verification of correct pK19mobsacB derivatives by PCR/sequencing |