TABLE 2.
Oligonucleotides used in this study
| Use and name | Sequence (5′-3′) | Construct produceda |
|---|---|---|
| RT-PCR | ||
| yceG RT1 F | GGGATGCGGCTGCAGACCGATCC | NA |
| g1 RT1 R | GTCAACCTCCTGCGGCGTC | NA |
| g15-16 RT2 F | GGGAGGTGTTTCAATTCGCC | NA |
| g15-16 RT2 R | GGTAAAGATCGGCCCAATGCG | NA |
| g16-cysE1 RT3 F | GCTGGCACAATGGTTGC | NA |
| g16-cysE1 RT3 R | GTGAATATTGCCCAGCA | NA |
| cyse1-1701 RT4 F | AGGACATGGCCTATTTCGTGC | NA |
| cyse1-1701 RT4 R | CATCACATGGTTCAGGTTCG | NA |
| qPCR | ||
| cysE1 qF | ATGCTGCATTCGGTGACA | NA |
| cysE1 qR | TGAATATTGCCCAGCACCTT | NA |
| cysE2 qF | CCAGCTTGACCAGGATGTC | NA |
| cysE2 qR | CGATGCGATGGTGGATCA | NA |
| uvrD qF | CAGAAGGAACACACGGTCAA | NA |
| uvrD qR | AAAGTGTCAGGCGGAATCTC | NA |
| Gene knockout constructs | ||
| Gent F | GATCCCCTGATTCCCTTTGT | pDS3, pDS4 and pDS5 |
| Gent R | CTTGAACGAATTGTTAGG | pDS3, pDS4 and pDS5 |
| cysE1 Up F | CGACTCTAGAGGATCCAGGTGTCGGATGTGTATGGC | pDS1 and pDS3 |
| cysE1 Up R | AACAATTCGTTCAAGCGGGTTTTGGACATGGGACT | pDS1 and pDS3 |
| cysE1 Down F | GGGAATCAGGGGATCCGGTCAACATGGATCAGCAG | pDS1 and pDS3 |
| cysE1 Down R | CGGTACCCGGGGATCCCAAAAACAGCCCGATCACCC | pDS1, pDS3, and pDS5 |
| cysE2 Up F | CGACTCTAGAGGATCCGATGCGCTGGCCCAGATCC | pDS6, pDS7, and pDS8 |
| cysE2 Up R-Gent | GGGAATCAGGGGATCACCGGAACCAGGGAAAACC | pDS7 |
| cysE2 Down F-Gent | AACAATTCGTTCAAGATGTCTGGCTGACCGAAAGC | pDS7 |
| cysE2 Down R | CGGTACCCGGGGATCTCGTCGTAATCGACGAAGG | pDS6, pDS7, and pDS8 |
| cysE2 Up R-Kan | GACACATGCAGCTCCACCGGAACCAGGGAAAACC | pDS8 |
| cysE2 Down F-Kan | ATTTTGAGACACAACATGTCTGGCTGACCGAAAGC | pDS8 |
| g16 Up F | CGACTCTAGAGGATCCGGGAGGTGTTTCAATTCGCC | pDS2, pDS4, and pDS5 |
| g16 Up R | AACAATTCGTTCAAGGTAAAGATCGGCCCAATGCG | pDS2 and pDS4 |
| g16 Down F | GGGAATCAGGGGATCCGACGATCTTGCAGAGGTGA | pDS2 and pDS4 |
| g16 Down R | CGGTACCCGGGGATCCCATCACATGGTTCAGGTTCG | pDS2 and pDS4 |
| Overexpression constructs | ||
| pPuf F | CGACTCTAGAGGATCCGAGCTTCGGAATCTGCG | pCMF151 and pCMF152 |
| pPuf R | CATAACAACCTCCGGATTGGCAGAC | pCMF151 and pCMF152 |
| cysE1 OX F | CCGGAGGTTGTTATGTCCAAAACCCGTGCG | pCMF152 |
| cysE1 OX R | CGGTACCCGGGGATCGTCAGCCCTCGCAGTCGC | pCMF152 |
| g16 OX F | CCGGAGGTTGTTATGCGGCCGATCCTGATC | pCMF151 |
| g16 OX R | CGGTACCCGGGGATCTCACAGATCCGCATCCAC | pCMF151 |
| puf con F | CGACTCTAGAGGATCCGAGCTTCGGAATCTGCG | pCMF278 |
| puf con R | CGGTACCCGGGGATCCATAACAACCTCCGGATT | pCMF278 |
| Reporter construct | ||
| pGTA F | CGACTCTAGAGGATCGATTGTCGATCAGATCAC | pCMF130 |
| pGTA R | GCTGACCATCGCCAGGGCCAGTTCC | pCMF130 |
| mCherryRc F | CCTGGCGATGGTCAGCAAGGGGGAG | pCMF130 |
| mCherryRc R | CGGTACCCGGGGATCCTGTTGTGTGGAATTGTGAGC | pCMF130 |
| Protein purification | ||
| H6-Cyse1 F | TCCAGGGACCAGCAATGATGTCCAAAACCCGTGCG | pCMF140 |
| H6-CysE1 R | TGAGGAGAAGGCGCGTCAGCCCTCGCAGTCGCAGC | pCMF140 |
| H6-Cyse2 F | TCCAGGGACCAGCAATGACGCTTGACACGCAACG | pDS9 |
| H6-CysE2 R | TGAGGAGAAGGCGCGTCGGTCATTCACGTTCTCC | pDS9 |
a
NA, not applicable.