TABLE 2.
Primers used in creating mutationsa
Mutation created | Primer | Position of 3′ end of primer | Sequence of primer (5′ → 3′) |
---|---|---|---|
1 | −320 | GCAGAATTCCCCCATCAGTGGATG | |
4 | —b | CCAGGGTTTTCCCAGTCACGACG | |
+2 | 2 | −172 | CCACAACTTAGCTAGCAATTTCTC |
3 | −151 | GAGAAATTGCTAGCTAAGTTGTGG | |
+4 | 5 | −172 | CCACAACTTGCTAGCAGCAATTTCTC |
6 | −151 | GAGAAATTGCTGCTAGCAAGTTGTGG | |
+6 | 7 | −172 | CCACAACTTAGCTAGCCAGCAATTTCTC |
8 | −151 | GAGAAATTGCTGGCTAGCTAAGTTGTGG | |
+8 | 9 | −172 | CCACAACTTAAGCTAGCTCAGCAATTTCTC |
10 | −151 | GAGAAATTGCTGAGCTAGCTTAAGTTGTGG | |
+10 | 11 | −172 | CCACAACTTAAGCTAGATCTCAGCAATTTCTC |
12 | −151 | GAGAAATTGCTGAGATCTAGCTTAAGTTGTGG | |
+100 | 13 | —c | TAGAGAAATTGCTGGTTGTCCCAATTCTTGTTG |
14 | —c | GGCTGAATCCCACAACTTAGGGTAAGTTTTCCGTATG | |
15 | −142 | CATACGGAAAACTTACCCTAAGTTGTGGGATTCAGCC | |
16 | −174 | CAACAAGAATTGGGACAACCAGCAATTTCTCTA | |
Δ1&2 | 17 | −142 | CGAGAATTCTAAGTTGTGGGATTCAGCC |
Δ1&2, +100 | 18 | —c | CGAGAATTCGTTGTCCCAATTCTTGTT |
Inserted bases are shown in bold, and ensuing restriction sites are underlined. The numbering refers to the startpoint of transcription of the ilvIH operon (20).
This primer corresponds to sequences in lacZ in plasmid pCV156. Plasmid pCV156 contains the region from −332 to +29 of the ilvIH operon, followed by trp and lac sequences.
These primers correspond to sequences in the gfp gene, encoding green fluorescent protein.