Table 2 Primers used in this study
From: Role of the gerA operon in L-alanine germination of Bacillus licheniformisspores
primer name | sequencea | Application |
|---|---|---|
Primers used in the construction/verification of the disruption mutant (NVH-1307) | ||
Upper ΔgerAA F BamHI | 5´- AATCGGATCCCAAGGAACACATCCATGAA-3´ | Amplification of the upper fragment of pMAD_SpRΔgerAAb |
Upper ΔgerAA R BamHI | 5´- TCAACAAAAATTGGGATCCGTCCATTAAA-3´ | Amplification of the upper fragment of pMAD_SpRΔgerAAb |
Lower ΔgerAA F EcoRI | 5´- TCTTCACCGAATTCGCTAGGCAAAGAA-3´' | Amplification of the lower fragment of pMAD_SpRΔgerAAb |
Lower ΔgerAA R EcoRI | 5´- AAATGGAATTCACCGTCAAAGCTCTG-3´ | Amplification of the lower fragment of pMAD_SpRΔgerAAb |
Upper ΔgerAA F2 | 5´- TGAAAATTTCGCCAAACACT-3´ | Verification/sequencing of NVH-1307 b |
specR R | 5´- TGATATGATCTTTCATTTCCATAAAAC-3´ | Verification/sequencing of NVH-1307 c |
Lower ΔgerAA R2 | 5´- TTCGGCAGAAACATCATCAG-3´ | Verification/sequencing of NVH-1307 b |
specR F | 5´- ATTGAATGGACTAATGAAAATGTAAA-3´ | Verification/sequencing of NVH-1307 c |
Primers used in the construction/verification of the complementation plasmid (pHT315gerA_MW3) | ||
pHT315gerAwt_SalI F1 | 5´-CAATCTGTCGACGTTTCCCCGTAAGCCTGATT-3´ | Amplification of the gerA fragmentb,d |
pHT315gerAwt_XbaI R1 | 5´-GTGAGGTCTAGACCGATCGTGAAGAAAAGCAT-3´ | Amplification of the gerA fragmentb,d |
ASF | 5´-AAAGAAGCCTTGGAGAAGTGA-3´ | Verification/sequencing of pHT315gerA_MW3b |
AOR | 5´-CGCTTTGCCCTGGATATAGA-3´ | Verification/sequencing of pHT315gerA_MW3b |
4AF | 5´-CAATCCGCTAGGCCAGAC-3´ | Verification/sequencing of pHT315gerA_MW3b |
6AF | 5´-GCGGACTGAGCCTGAATATG-3´ | Verification/sequencing of pHT315gerA_MW3 b |
8AF | 5´-CGCTCAGGATCCGTCTAAAG-3´ | Verification/sequencing of pHT315gerA_MW3b |
A9F 2/8/15 | 5´-CAGATCGAAGCGCTGAATTT-3´ | Verification/sequencing of pHT315gerA_MW3 b |
pHT315R | 5´-GGAGAAAATACCGCATCAGG-3´ | Verification/sequencing of pHT315gerA_MW3e |