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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
  1. aThe restriction sites are underlined and may differ in sequence from the template genome of Veith et. al.[48].
  2. bThe primer is complement to the genome of Veith et. al.[48].
  3. cThe primer is complement to the SpR cassette of pMAD_SpR.
  4. dThe gerA fragment amplified contains a sequence 151 bp upstream of gerAA, gerAA, gerAB, gerAC and 177 bp downstream of gerAC relative to the genome of Veith et. al.[48].
  5. eThe primer is complement to the pHT315 vector.