Table 2.
Primers and probe used in this study.
| Primer designation | Sequencea,b | Purpose |
|---|---|---|
| 5′ BtFlaB | CTGGAATGGGTGTTGCAGGA | RT-PCR; PCR Screening |
| 3′ BtFlaB | CTCCCTCTTGTTGTGCACCT | RT-PCR; PCR Screening |
| 5′ Chrom | GATTATCAATCATAATACATCAGC | RT-PCR |
| 3′ Chrom | TCTAAGCAATGACAAACATATTGG | RT-PCR |
| 5′bt0790-790A link RT-PCR (P1—Figure 1) | CCCTGTTCGTTATGAAAATGCTTTGCTTGG | RT-PCR |
| 3′bt0790-790A link RT-PCR (P2—Figure 1) | GCAAAGAAACTCGCAAGCATTGGGTCC | RT-PCR |
| 5′bt0790A-791 link RT-PCR (P3—Figure 1) | GCTCCTAATTCTCCTGCAGATATTGGG | RT-PCR |
| 3′bt0790A-791 link RT-PCR (P4—Figure 1) | GGCCTACATCAAAAGAATCACCTGC | RT-PCR |
| 5′bt0790A ORF RT-PCR (P3—Figure 2) | TGTAAGACTTCCAAGAGGCAAGGG | RT-PCR; PCR Screening |
| 3′bt0790A ORF RT-PCR (P4—Figure 2) | TTTCATTTACCACAGGTCCACCGG | RT-PCR; PCR Screening |
| 5′bt0790 ORF RT-PCR | GGGAAGTTTTAGTAAAGTGTTAGCCG | RT-PCR |
| 3′bt0790 ORF RT-PCR | ACTGCGGCATTATTTTTGTCATCTACA | RT-PCR |
| 5′bt0791 ORF RT-PCR_v2 | CCATTACCAAGGGACGTAGAAA | RT-PCR |
| 3′bt0791 ORF RT-PCR_v2 | ACAATCTCATCACCACCAACA | RT-PCR |
| 5′bb0790-bb0791 link (P5—Figure 1) | GGCTGCAATTTTAATAGTTTTGGGATC | RT-PCR |
| 3′bb0790-bb0791 link (P6—Figure 1) | CGTTCATCATAAAAACATGCCTCATC | RT-PCR |
| bt0790 IDT SYBR FWD | ACTTGATTTACATGAGACTTGAAGC | qRT-PCR |
| bt0790 IDT SYBR REV | AAAGAGTCGGCTAACACTTTACT | qRT-PCR |
| bt0790A IDT SYBR FWD | CCAATGCTTGCGAGTTTCTTT | qRT-PCR |
| bt0790A IDT SYBR REV | CCCATATCCACCCAAACTGTTA | qRT-PCR |
| bt0791 IDT SYBR FWD | CCATTACCAAGGGACGTAGAAAC | qRT-PCR |
| bt0791 IDT SYBR REV | TCACTTCCACCGCCTCTATAA | qRT-PCR |
| 5′ BtflaB SYBR/ABI | AAAAACAGCTGAAGAGCTTGGAAT | qRT-PCR |
| 3′ BtflaB SYBR/ABI | CACCCACATGTACTCTTAATGTCCAT | qRT-PCR |
| 5′ F1 bt0790A KO | CACCTGATGAAGCTTATATGTTTTTTA | Mutagenesis Cloning |
| 3′ F1 bt0790A KO_AscI | GGCGCGCCCTCTACTCTCAGCAAGCATCATACC | Mutagenesis Cloning |
| 5′ F2 bt0790A KO_AscI | GGCGCGCCAATATTATTGAGAGTATTATAGAG | Mutagenesis Cloning |
| 3′ F2 bt0790A KO_BssHII | GCGCGCTCCCTACCAACAAGATAATGATGCC | Mutagenesis Cloning |
| 3′ F1 bt0790A KO_NdeI | CATATGCTCTCAAAAGCTAATTAATGTTATGATAG | Mutagenesis Cloning |
| 5′ F2 bt0790A KO_AscI_v2 | GGCGCGCCGCTAAGGTCTTTGCAAATGGTCTTGGTG | Mutagenesis Cloning |
| 5′ BtflgB-AscI | GGCGCGCCAGCACCCGGTAGCAAGTTAAAAAAATTTG | Mutagenesis Cloning |
| 3′ Gent-AscI | GGCGCGCCTTAGGTGGCGGTACTTGGGTCG | Mutagenesis Cloning |
| 5′ PromLess Bt Gent | GGCGCGCCATAGAGGGTCATATGTTACGCAGCAGC | Mutagenesis Cloning |
| 5′aacC1 diag | GCAACGATGTTACGCAGCAG | PCR Screening |
| 3′aacC1 diag | GCATCACTTCTTCCCGTATGC | PCR Screening |
| 5′ Bt0790A ext diag_V2 (P1—Figure 2) | GATAAAGGAGTTTTGAAAGTTAAGAAAG | PCR Screening |
| 3′ Bt0790A ext diag_V2 (P2—Figure 2) | CATAAAGCAATAAGACAAACACTCTCT | PCR Screening |
| BtflaB F | CCAGCATCATTAGCTGGATCAC | qPCR |
| BtflaB R | GTTGTGCACCTTCCTGAGC | qPCR |
| BtflaB-Probe | /5YakYel/TGCAGGTGA/ZEN/AGGTGCGCAGGTT/3IABkFQ/ | qPCR |
a
Relevant restriction sites are indicated by bold lettering.
b
YakYel, 5′ Yakima Yellow dye; ZEN, ZEN internal quencher; IABkFQ, Iowa Black FQ 3′ quencher.