TABLE 3.
Major primers used in this studya
| Primer | DNA sequence (5′–3′) | Product size (bp)b | Target gene and function |
|---|---|---|---|
| ChuB_F | GATGATTTTTGGTGGGATTTTT | 2,063 | Amplify chuB fragment |
| ChuB_R | TTTAAATCAAGGGCTGAAGTGG | ||
| Erm_PacI_F | CCCTTAATTAAAGCTTTGGCTAACACACACG | 1,140 | Erythromycin resistance cassette |
| Erm_PacI_R | CCCTTAATTAAAATAGGTACACGAAAAACAAGTTAAGG | ||
| CjeI_F | GGAGCAAAACGAAGAATTGC | 4,627 | CjeI fragment from C. jejuni NCTC 11168 |
| CjeI_R | CAGAGCAAACCTCCATCACA | ||
| CjeI_F1 | GCAATCCGCAAAATGAAACT | 418 | Detect insertional mutation |
| CjeI_R1 | TTTGCATATCTTGCCACTCC | ||
| Cm_PacI_F | CCCTTAATTAATGCTCGGCGGTGTTCCTTT | 801 | Chloramphenicol resistance cassette |
| Cm_PacI_R | CCCTTAATTAAGCGCCCTTTAGTTCCTAAAG | ||
| Cm_F (MfeI) | GAGCAATTGTGCTCGGCGGTGTTCCTTT | 801 | Chloramphenicol resistance cassette |
| Cm_R (MfeI) | GAGCAATTGGCGCCCTTTAGTTCCTAAAG | ||
| CjeI_F2 | AAAAAGAATCAAACATTACCATCTTTC | 5,539 (NCTC 11168) | Detect gene replacement in transformants |
| CjeI_R2 | AAAAAGCTTTATGGCGAGGA | 3,471 (CG8486) | |
| rrsF (NdeI) | GGAATTCCATATGCTGGAACTCAACTGACGCTAAG | 1,900 | rRNA gene |
| rrsR (SphI) | ACATGCATGCCTCTTGCACATTGCAGTCCTAC | ||
| pCjR_MCS_F1 | CTAGAGAATTCGAGCTCGGTACCGGATCCGTCGACCTGCAGAGCGCTC | 42 | Introduce MCS sites into pCjR |
| pCjR_MCS_R1 | AATTGAGCGCTCTGCAGGTCGACGGATCCGGTACCGAGCTCGAATTCT | ||
| Cj0299_pMW10_F (XbaI) | TGCTCTAGAACTTTTTCTCATTTTGCATACCTCA | 280 | Cj0299 overexpression promoter |
| Cj0299_pMW10_R1 (KpnI) | CGGGGTACCGCCCAAACACTAAATTTAAGAAAA | ||
| CjeI_F4 (KpnI) | CGGGGTACCATCAATCAAGGTGGCTTTGG | 4,146 | Cloning whole ORF of CjeI |
| CjeI_R4 (BamHI) | CGCGGATCCAACCCGCTCCGCTTAATATC | ||
| rrs_Seq_F | TCACTCGAAGCCGGAATACT | 231 | Confirm cloning within the ribosomal site |
| rrs_Seq_R | CAGCTGAGCTAATTCCCCATA | ||
| CjeI_F5 (NdeI) | GGAATTCCATATGATGATTACAAAAGATAATTTAAAACAAGTTC | 4,017 | Generate rCjeI expression construct |
| CjeI_R5 (SalI) | GCGCATGTCGACAGAAAATAAATATTTTTGCAAGATTTTTTC | ||
| CjeI_F_Seq | AACATAGGTTTTGGTAGCACAGAA | Detect cloning of CjeI in suicidal vector | |
| Cm-Seq_R | CAGGGCGTATTGCCAAAATA | Detect cat insertion in CjeI mutant |
a
Restriction sites are underlined in the primer sequence and the names are identified in parentheses.
b
Size of amplicon in different strains. The strain name is in parentheses.