From: Cell factory for γ-aminobutyric acid (GABA) production using Bifidobacterium adolescentis
Primer | Sequence (5’—> 3’) | Template | Purpose |
---|---|---|---|
gadBC_OP_Fw | ccagctcttcgACAacctgcccatcgtagc | B. adolescentis 4–2 genomic DNA | Amplify gadBC gene with the original promoter |
gadBC_OP_RV | ccagctcttcgCTAtcagtattccggattcactagc | ||
pKKT427 Fw | caagctcttcgTAGgccaccgtcgccaagg | pKKT427 | Amplify pKKT427 plasmid backbone |
pKKT427 Rv | caagctcttcgTGTgcctgcatgcaagctt | ||
gadBC Fw | ccagctcttcgatgtcagaaacacattccacc | B. adolescentis 4–2 genomic DNA | Amplify gadBC gene without the original promoter |
gadBC Rv | caagctcttcgtcagtattccggattcactagc | ||
pKKT427_ter_Fw | ccagctcttcgTGActgactcactgaacgg | pBCMAT_Pgap_TdppA2 | Amplify pKKT427 plasmid backbone, including the gap promoter and terminator |
pKKT427_pro_rv | ccagctcttcgCATgatgttctccttgggtca | ||
gadB_RT1_Fw | catgttcctgcgtttgggat | B. adolescentis cDNA | gadB quantitative expression |
gadB_RT1_Rv | ccgtcgttccacagcgta | ||
gadC_RT1_Fw | cgtcggtttcgtcgctt | gadC quantitative expression | |
gadC_RT1_Rv | cacaagaatcgcatatgaaacgcta | ||
16srRNA_FW | cacattccaccgttacacc | Normalize gene expression in B. adolescentis | |
16srRNA_FW | cgttatccggaattattggg |