Table 2.
Primers used in the present study.
| Primer | Sequence 5′-3′ | Amplified DNA |
|---|---|---|
| Phusion PCR for carB disruption | ||
| carBcrispfuz1 | CTGTGCAGGTGCCAGCGCATCACC | carB promoter and coding sequence upstream from protospacer |
| carBcrispfuz2 | GGTGGTCTTGAATGCGCTCGTCCAG | |
| carBcrispfuz3 | TGGAGCTGCTGCGATGCGACAAC | carB coding sequence downstream from protospacer |
| carBcrispfuz4 | GTCTTGCTCTTCATATGACCAATAG | |
| carBcrispfuz5 | CCGATCTGGACGAGCGCATTCAAGACCACCTGCCTCAGCATTGGTACTTG | pyrG with own promoter and terminator sequences |
| carBcrispfuz6 | GTAGTTGTTGTCGCATCGCAGCAGCTCCAGTACACTGGCCATGCTATCG | |
| carBcrispfuz7 | CTTGTGGTAGACAATGTAGTTGTC | phusion pcr product |
| carBcrispfuz8 | CGTCAAAGCTCTCCTGATAAGCCTC | |
| Primers for analysis of the carB mutants | ||
| MccarRPfw | ATGCTGCTCACCTACATGGAAG | carRP |
| MccarRPrev2 | TTAAATGGTATTTAGATTTCTCA | |
| MccarB1 | ATGTCCAAGAAACACATTGTCATTA | carB |
| MccarB2 | TTAAATGACATTAGAGTTATGAACG | |
| MccarBupfw | TAGCCAATGACAGCGGTGACGC | upstream region from protospacer |
| MccarBuprev | GTTGTCGCATCGCAGCAGCTCCA | |
| MccarBdwfw | TGGAGCTGCTGCGATGCGACAAC | downstream region from protospacer |
| MccarBdwrev | CGTCAAAGCTCTCCTGATAAGCCTC | |
| Phusion PCR for hmgR 2 disruption | ||
| h2p1 | ACGGAGCAAGTTTACATTCATACC | hmgR2 promoter and coding region upstream from protospacer |
| h2p2 | GATTGGCGGTCTATCATATTTCAG | |
| h2p3 | CTCAACATCTCTTGTACTATGCCC | hmgR2 coding sequence downstream from protospacer |
| h2p4 | ATACGTTTGTACCGATGAAGGGA | |
| h2pyrG5 | CTCATTGCTGAAATATGATAGACCGCCAATCGTACAATTCCCTGCCTTCTGGAAG | pyrG with own promoter and terminator seqences |
| h2pyrG6 | CGATACAGGGCATAGTACAAGAGATGTTGAGCGCGTGAAAGACCTGCCTTGACTC | |
| h2p7 | TGTGAAATTGGCCTATAATCAGCCT | phusion pcr product |
| h2p8 | GGACAGACATTCCATCGTATACAG | |
| Primers for analysis of the hmgR 2 mutant | ||
| h2cdns1 | ATGTTGAAAAACGTCAAAAAAGAT | hmgR2 |
| h2cdns2 | CTATGATTTAATACAACTTCCA | |
| Primers used in the qPCR experiment | ||
| MCactinF | CACTCCTTCACTACCACCGCTGA | 117 bp of actin |
| MCactinR | GAGAGCAGAGGATTGAGCAGCAG | |
| H2_RT_F | CTCGTATCATCTGTGCCTCTG | 107 bp of hmgR2 |
| H2_RT_R | AGCAGTGTTACGGTTGTGAG | |