Copyright
©The Author(s) 2015.
World J Biol Chem. Aug 26, 2015; 6(3): 249-264
Published online Aug 26, 2015. doi: 10.4331/wjbc.v6.i3.249
Published online Aug 26, 2015. doi: 10.4331/wjbc.v6.i3.249
Table 1 Oligonucleotides used for amplifying the genes expressing Bacillus peroxiredoxins proteins and its Cys residue-mutated proteins
| Sequence of primer for point mutation | |
| AhpC_H1 | |
| C37S F | AGCGTATTCTCTTTCTACCCA |
| C37S R | TGGGTAGAAAGAGAATACGCT |
| C47S F | TTCTCTTTCGTATCTCCAACTGAGCTT |
| C47S R | AAGCTCAGTTGGAGATACGAAAGAGAA |
| C166S F | CCAGGTGAAGTTTCTCCGGCTAAATGG |
| C166S R | CCATTTAGCCGGAGAAACTTCACCTGG |
| AhpC_H2 | |
| C52S F | TTCACTTTTGTTTCTCCGACAGAAATT |
| C52S R | AATTTCTGTCGGAGAAACAAAAGTGAA |
| C169S F | ACTGGCGGACTCTCTCCGGCTAACTGG |
| C169S R | CCAGTTAGCCGGAGAGAGTCCGCCAGT |
| Sequence of primer for protein expression | |
| AhpC_H1 | |
| Forward | CCGCATATGTCTTTAATCGGTAAAGAAGATC |
| Reverse | CCGGTCGACTTAGATTTTACCTACTAGATCAAGG |
| AhpC_H2 | |
| Forward | CCGCATATGGCAGAACGTATGGTAGGTAA |
| Reverse | CCGGTCGACTTAAAGTGTTTTTTGGCCTGGTTT |
| B_BCP | |
| Forward | CCGCATATGACTATAGAAATCGGACAAAAAC |
| Reverse | CCGGTCGACTTACTTTTCAGACATTTTGAGG |
| B_TPx | |
| Forward | CCGCATATGGCTGAAATTACATTCAAAGGC |
| Reverse | CCGGTCGACTTACTTTCTAATGCAGCAGC |
Table 2 Oligonucleotides used for amplifying the DNA fusion construct to knockout Bacillus Prx genes
| Prx null mutants | Primer names | Sequences of paired degenerated primers (5’→3’) |
| ΔAhpC_H1 | H1_Up F | CCTTATTTCACAGATAAGCTCCA |
| Kan/H1 R’ | CCTATCACCTCAAATGGTTCGCTG AATGTATATTCCTCCTAAAAATGTAT | |
| Kan/H1 F’ | CGAGCGCCTACGAGGAATTTGTATGGAGTGCATTCAATTGGTACTTG | |
| H1_Down R | ACGTGCAGTGTAGATCGCTGC | |
| ΔAhpC_H2 | H2_Up F | AAGCTGCGGATTTGAGTTGTCC |
| Kan/H2 R’ | CCTATCACCTCAAATGGTTCGCTGATGTATCCCTCCAATTTATTGTTTG | |
| Kan/H2 F’ | CGAGCGCCTACGAGGAATTTGTAT TTCTTTCCAAGAACGAAAAGCGG | |
| H2_Down R | ATCTAAAATTGAGGAAAAACATCAA | |
| ΔB_BCP | BCP_Up F | AATGGTACTGTGATCACTTCGTTT |
| Kan/BCP R’ | CCTATCACCTCAAATGGTTCGCTG TACGTTACCTCCGGATGTTTTTTT | |
| Kan/BCP F’ | CGAGCGCCTACGAGGAATTTGTAT ATCTCTATGAGCCTATGCTTACTT | |
| BCP_Down R | AGTAGTCGATATGTGCATGCATT | |
| ΔB_TPx | TPx_Up F | TTTTTCGCTTTGGACATGCTAT |
| Kan/TPx R’ | CCTATCACCTCAAATGGTTCGCTGTATAATTCCTCCCTTTTGTATGTAT | |
| Kan/TPx F’ | CGA GCGCCTACGAGGAATTTGTAT GCAGGGAAAAAAGCTCCAGGC | |
| TPx_Down R | CGTCAGCTTTGAGCTCAAACG |
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Citation: Cha MK, Bae YJ, Kim KJ, Park BJ, Kim IH. Characterization of two alkyl hydroperoxide reductase C homologs alkyl hydroperoxide reductase C_H1 and alkyl hydroperoxide reductase C_H2 in
Bacillus subtilis . World J Biol Chem 2015; 6(3): 249-264 - URL: https://www.wjgnet.com/1949-8454/full/v6/i3/249.htm
- DOI: https://dx.doi.org/10.4331/wjbc.v6.i3.249