Table 3 Oligonucleotides used in this study
Primers | Sequence | Gene region | Size (bp) | Reference |
|---|---|---|---|---|
UP 378 | GCGTTTTGACCATCTTCGT | Â | Â | Â |
LP 378 | ACAGGAGCAGTTTCAGACAG | 378 bp fragment stx 2 -A | 378 | 49 |
S2Aup | ATGAAGTGTATATTATTTA | Binds stx2-A subunit at initial codon | - | 7 |
GK4 | TCAGTCATTATTAAACTG | Binds stx2-B subunit at final codon | - | 50 |
Cm 5 | TGTGTAGGCTGGAGCTGCTTC | Â | Â | Â |
Cm-3 | CATATGAATATCCTCCTTAG | cat gene | 1015 | This study |
Cm 5-stx | GAAGCAGCTCCAGCCTACACA ACGAAGATGGTCAAAACGCG | Â | Â | Â |
Cm3-stx | CTAAGGAGGATATTCATATG AGGAGTTAAGTATGAAGAAG | Used for construction of the 5' fragment and 3' fragment of the cat gene | - | This study |
Tc 5 | TCAGCCCCATACGATATAAG | Â | Â | Â |
Tc-3 | TGGAGTGGTGAATCCGTTAG | tet gene | 1180 | This study |
Tc 5-stx | CTTATATCGTATGGGGCTGA ACGAAGATGGTCAAAACGCG | Â | Â | Â |
Tc3-stx | CTAACGGATTCACCACTCCA AGGAGTTAAGTATGAAGAAG | Used for construction of the 5' fragment and 3' fragment of the tet gene | - | This study |
Tc-int | TGTCGGAATGGACGATAT | Bind Tc cassette at 100 bp of 5' of tet | - | This study |
Rho | ATATCTGCGCCGGGTCTG | Binds rho independent terminator sequence upstream stx 2 -A subunit | - | This study |
RR46 LP | GAGCTCTAAGGAGGTTAT | Â | Â | Â |
RR46-UP | GTGCAGTACTCATTCGTT | Red recombinase gene in pKD46 vector | 457 | This study |
pBC KS | TCGAGGTCGACGGTATC | Â | Â | Â |
M13 rev | GGAAACAGCTATGACCATG | Detection of pBC-SK+ vector | 158 | Stratagene |