TABLE 2.
Primers and oligonucleotide sequences
| Primera | Oligonucleotide sequenceb | Genec |
|---|---|---|
| A | 5′-CTCTCTGCTAGCAGGAGGAATTCACCATGAATACCATTTTTTCCGC-3′ | dacA (F) |
| B | 5′-GCTTTCAAAACGCACTAACGG*GTTACCTTCCGGGATTTCTTG-3′ | dacA (R) |
| C | 5′-CCGTTAGTGCGTTTTGAAAGCCG-3′ | dacB (F) |
| D | 5′-CTCTCTCTCCAAGCTTCTAATTGTTCTGATAAATATCTTTATAC-3′ | dacB (R) |
| E | 5′-CTCTTCCACATTTTCCATCAC*AACCAGCGGGCGTTGCTCG-3′ | dacA (R) |
| F | 5′-GTGATGGAAAATGTGGAAGAGGGCGG-3′ | dacC (F) |
| G | 5′-CTCTCTAAGCTTTTAAGAGAACCAGCTGCC-3′ | dacC (R) |
| H | 5′-CCCGACAGATTCCAGGGT*AACCAGCGGGCGTTGCTCG-3′ | dacA (R) |
| I | 5′-ACCCTGGAATCTGTCGGGGAAGGCAG-3′ | dacD (F) |
| J | 5′-CTCTCTAAGCTTTCAGGCCTTATGGTGGAAATAATC-3′ | dacD (R) |
| K | 5′-CAGACGACCGATACCGCGAAG*GTTACCTTCCGGGATTTC-3′ | dacA (R) |
| L | 5′-CTTCGCGGTATCGGTCGTCTGG-3′ | proW (F) |
| M | 5′-CTCTCTCTCAAGCTTTTACTTAATGAATGGGCGGGTC-3′ | proW (R) |
| N | 5′-GGCATCAGGTTTAATTGGCGTCAC*GGTTTCAAAGAAACGGAAGCC-3′ | dacA (R) |
| O | 5′-GTGACGCCAATTAAACCTGATGCC-3′ | dacC (F) |
| P | 5′-CTCTTTGCTAGCAGGAGGAATTCACATGACGCAATACTCCTCTC-3′ | dacC (F) |
| Q | 5′-CTTTACCTACTTTCAGTGGGTTAAC*GGTTTCAAAGAAGCGGAAACCCCAGGT-3′ | dacC (R) |
| R | 5′-GTTAACCCACTGAAAGTAGG-3′ | dacA (F) |
| S | 5′-GCATGCAAGCTTCTAGATTTTTAACCAAACCAGTGATG-3′ | dacA (R) |
a
Primer designation in Table 1.
b
Underlined sequences are complementary to the 5′ (forward primer) or 3′ (reverse primer) ends of the gene fragments in each fusion construct. Italicized sequences are the portions of primer P2 that are complementary to the 5′ end of the gene fragment (primer P3) that will be fused to the 3′ end of the hybrid gene. An asterisk within the sequences of oligonucleotides used as P2 primers designates the fusion site between two coding sequences. Sequences in bold designate HindIII (AAGCTT) and NheI (GCTAGC) sites.
c
Gene to which each oligonucleotide anneals in PCR amplification. F, forward primer (P1 or P3 in the text); R, reverse primer (P2 or P4 in the text).