TABLE 1.
Plasmids used in two-hybrid assays
| Plasmid | Fused genesa | Vector | PCR primersb used or cloning strategy |
|---|---|---|---|
| pHA409 | gal4AD::lctAc | pGAD424 | LCTA1, ATTAAGAATTCATAATGAAAGAACAAAACTCT (EcoRI) |
| LCTT4, ATATTCTGCAGCGATACGTAACTTTTTAT (PstI) | |||
| pHA689 | gal4AD::lctM | pGAD424 | LCTM1, TAATAGAATTCATAGTGAAAAAAAAGACTTAC (EcoRI) |
| LCTM2, TTATACTGCAGATATTAATCAACATATGGC (PstI) | |||
| pHA888 | gal4AD::lctM | pGAD424 | The 2.1-kb NdeI fragment internal to the pHA689 insert was replaced by the corresponding fragment from the cloned lacticin 481 operon |
| pHB246 | gal4BD::lctAc | pGBT9 | LCTA1 and LCTT4 |
| pHB685 | gal4BD::lctM | pGBT9 | LCTM1 and LCTM2 |
| pHB887 | gal4BD::lctM | pGBT9 | The 2.1-kb NdeI fragment internal to the pHB685 insert was replaced by the corresponding fragment from the cloned lacticin 481 operon |
a
The junctions between the vectors and the 5′ ends of the inserts have been verified by sequencing. The inserts carrying lctA were entirely sequenced.
b
The names of the primers and their 5′→3′ sequences are given. Only boldfaced bases are complementary to the target sequence. Restriction sites are underlined in the sequences and identified in parentheses.
c
The insert is a 0.34-kb DNA fragment resulting from the EcoRI-BclI digestion of the larger fragment amplified with LCTA1–LCTT4.