Table 2.
Primers used in this study.
| Primers | Sequence (5'-3') | Purpose |
|---|---|---|
| Plasmid construction | ||
| Int48-F | CCGGAATTCATGGGTAGTATTAACTCTCG | pInt48 |
| Int48-R | CGCGGATCCTTATCCTCTTAGTTTTTGGTTC | |
| Xis21-F | CCGGAATTCATGAACCCATCAAATCACG | pXis21 |
| Xis21-R | CGCGGATCCTAATTGATACTTTCGCGG | |
| Int2894-F | CCGGAATTCTTGTCTAAGGACTCGACGGAG | pInt2894 |
| Int2894-R | CGCGGATCCTTATTTGTTGTTCATCATCATTATTCC | |
| XisPO1-F | CCGGAATTCGTGAACATGAACCCATCAAATC | pXisPO1 |
| XisPO1-R | CGCGGATCCCTAATTGATACTTTCGCGGTTGG | |
| XisANA3-F | CCGGAATTCGTGAGCATGAACTCATTAAATAAAC | pXisANA3 |
| XisANA3-R | CGCGGATCCTTAGTACTTCCCATCTTCGACTG | |
| hipA-SalI-F | ACGCGTCGACGAACAGTTGACCATTCAGGC | pHipA |
| hipA-PstI-R | TGCACTGCAGTCATACCAATCCCCAACGCG | |
| hipB-SalI-F | ACGCGTCGACAGTGATAAACAAACGACTAC | pHipB |
| hipB-PstI-R | TGCACTGCAGTCATAAAAGCCATGTGACAC | |
| hipA-SalI-F | ACGCGTCGACGAACAGTTGACCATTCAGGC | pHipAB |
| hipB-PstI-R | TGCACTGCAGTCATAAAAGCCATGTGACAC | |
| pHGR01-PhipA-F | CCGGAATTCACTGTAGCGCATATTTAATAA | pHGR01-PhipA |
| pHGR01-PhipA-R | CGCGGATCCgtaatcatggTCATGAAAGCTCCCAAAGACATTATG | |
| pMD19-T-hipAB-F | TCATAAAAGCCATGTGACAC | pMD19-T-hipAB |
| pMD19-T-hipAB-R | GTCACCACATTAGTCCCACT | |
| Construction of ΔhipAB | ||
| hipAB-up-F | ACATGCATGCGAGATGAAACGCTTCAACTCG | pK18Cm-hipAB |
| hipAB-up-R | CCGGAATTCCAGTGGATAGCATTGACCAAC | |
| hipAB-down-F | CCGGAATTCGCTCGTAATCTAACGAGGTAAG | |
| hipAB-down-R | AGCGTCGACCCAGGTTACTAATTCTAGTCAC | |
| hipAB-wF | GTTTACATAAACCAGCAGCAC | Confirmation of ΔhipAB |
| hipAB-wR | GTCCATATTACTCACCTTAGC | |
| Construction of ΔhipAB Pint::lacZ and CN32 Pint::lacZ | ||
| pHGI01-Pint-F | CCGGAATTCAACGTCGAATGACGTTTTTAGCG | pHGI01-Pint |
| pHGI01-Pint-R | CGCGGATCCgtaatcatggGTAGTTAAGTCCAAAATGGTGAC | |
| mob-F | CAGAGCAGGATTCCCGTTGAGCA | Confirmation of ΔhipAB Pint::lacZ and CN32 Pint::lacZ |
| LacZ-R | TATTACGCCAGCTGGCGAAAGG | |
| Int-F | ATGATTAAGTGTCACTTTTCAAGG | |
| Int-R | CATTTGGCTGCGATTAGCTC | |
| Primers used in determination of the excision and circled form of CGI48 and GI21 | ||
| 21F | CCAAAGCGAGGTAAGACGT | ΔGI21 |
| 21R | TCGGAGACAGCGATGTATCG | |
| 21cirF | AGTGGGACTAATGTGGTGACTAGAATT | The circled GI21 |
| 21cirR | TGCAAGTGCATGGTTTTATGATG | |
| 48F | CCAAGTGAACGTTTATGATCGC | ΔCGI48 |
| 48R | GGTGTGTTTTTCATCGTTATGC | |
| 48cirF | CGAGAGTCTATTCGTAGAGAC | The circled CGI48 |
| 48cirR | AGAATATGGTCTAACCAAGC | |
| oF | CCGGAATTCATGATTAAGTGTCACTTTTCAAGG | cro/cI gene |
| oR | CGCGGATCCTTAGTCTGTACCTTGGATTTC | |
| Primers used in qPCR for CGI48 and GI21 in CN32 | ||
| q48F | GGCTCGCATATTTCTGTGCAA | Determine the excision rate of CGI48 |
| q48R | CCTTTGAGAGTGCTTTTAGCATAATG | |
| q21F | TTGGCGAGTTGCTCGAAATC | Determine the excision rate of GI21 |
| q21R | GGAACTGGGATGTGTTTTATTGC | |
| q48cF | CGAGAGTCTATTCGTAGAGAC | Determine the circular form of CGI48 |
| q48cR | AGAATATGGTCTAACCAAGC | |
| q21cF | AGTGGGACTAATGTGGTGACTAGAATT | Determine the circular form of GI21 |
| q21cR | TGCAAGTGCATGGTTTTATGATG | |
| CN32gyrB-qF | TTCGTACTTTGCTGTTGACCTTCT | Reference gene |
| CN32gyrB-qR | CTACGGTGCCATCCAATGCT | |
| Primers used in qPCR for GISpuPO1 in W3-18-1 | ||
| GISpuPO1-qF | AGGTCGCCGTCTCGATTTTA | Determine the excision rate of GISpuPO1 |
| GISpuPO1-qR | TGAGTCGGAAACATCATTAGACGTT | |
| W3181gyrB-qF | GCTCAGCCGCCTTTGTTTAA | Reference gene |
| W3181gyrB-qR | CGGCTCACCCGACATACC | |
| Primers used in qPCR for GISspANA3 in ANA-3 | ||
| GISspANA3-qF | GTCGAGCTCAAAGTACTCATCGAA | Determine the excision rate of GISspANA3 |
| GISspANA3-qR | GCTACAGCAGAAGCTAATCTCATTACTC | |
| ANA3gyrB-qF | CTGGTGAGCCTGTGCTCGAT | Reference gene |
| ANA3gyrB-qR | CAAGCGCCGCACCTAACTTA | |
Restriction sites included in oligonucleotide sequences are underlined.