Table 2.
Oligonucleotides used in this studyb
| Genea | Application | Primer | Sequence (5′→3′) |
|---|---|---|---|
| HP0058 | Mutagenesis | 0058-L3 | GCTAACTAACAAGATCACCG |
| PCAT-0058-R2 | 1-CGCTATGAGTTGTTGCTACA | ||
| CAT-0058-L1 | 2-GCGCTTATGACTATACATGC | ||
| 0059-R1 | CGGTGATCTTGTTAGTTAGC | ||
| HP1142 | Mutagenesis | 1143-L1 | AAGCGACATGCGAGAGATTG |
| PCAT-1142-R1 | 1- CCACTGCTTACATTCTGCTG | ||
| CAT-1142-L1 | 2- GAAGATGGTCAATTAGTAGG | ||
| 1142-R2 | TATAATGAGCTTCATGACCG | ||
| HP0058 | Strep tag fusion | 58s-L2 | ATGGTAGGTCTCAGCGCTTGTTAAGAGAAAAAGAAAATCTCAATA |
| 58Strep-PCAT | GGCGGATTAACAAAAACCGGACTATTTTTCAAATTGCGGGTG | ||
| 59PCAT | TGGCAGGGCGGGGCGTAAATGGGA | ||
| 59_dw | GCTCTTGTTTAAGGCTATCC | ||
| HP0058 | Expression | 58_up | TCAGAATTCATGATGGGTGCTCATATTATAG |
| 58_Strepdw | TCACTGCTATTTTTCGAACTGCGGGTGGCTCCACCCGCCTGATCCCATATCCACGATAGT | ||
| HP1142 | Expression | pASK7-1142-L1 | ATGGTAGGTCTCAGCGCGTGAGCGTGAATAGTAATGGCAAT |
| pASK7-1142-R1 | ATGGTAGGTCTCATATCACTTCATTCTCATCATCATTTTATAATG | ||
| Pcat | catgene with promoter | CATS1 | TCCGGTTTTTGTTAATCCGCC |
| CATAS1 | TTACGCCCCGCCCTGCCA |
a
Gene numbers refer to the H. pylori26695 genome sequence (27).
b
The 5′ extensions used for the fusion of PCR products to the catgene by megaprimer PCR are labeled as follows: 1 (5′-GGCGGATTAACAAAAACCGGA), complementary to the 5′ region of the catgene with promoter; 2 (5′-TGGCAGGGCGGGGCGTAA), complementary to the 3′ end of the catgene. The BsaI restriction site used for the protein expression via the IBA system is underlined.