TABLE 1.
Primers
| Name | Sequencea | Target positions (nt) and restriction sites |
|---|---|---|
| 5′ 5-7 | 5′-ATGTAGATCTTTCTCAGCTATATCCTG-3′ | IKe (1181–1207), BglII, AluI |
| 5′ 7 | 5′-CGCTTTAAAGTACCGTTTACATCCTGC-3′ | IKe (1469–1495), DraI |
| 3′ 5-7,7 | 5′-ATTAGATCTTCACTAGTCATGCTCATT-3′ | IKe (1584–1558), BglII |
| 3′ ATG→CTG | 5′-ATTAGATCTTCACTAGTCAT/GGCTCAT/GTTAA-3′ | IKe (1584–1555), BglII |
| 3′ stop codon | 5′-ATTAGATCTTCACTAGTCATGCC/TC/TATTTAA-3′ | IKe (1584–1555), BglII |
| 3′ TGG | 5′-ATTAGATCTTCACTAGTCATGCCCATTTAA-3′ | IKe (1584–1555), BglII |
| 3′ CAG | 5′-ATTAGATCTTCTGTAGTCATGCTCATTTAA-3′ | IKe (1584–1555), BglII |
| 5′ SD mutant | 5′-CTCAGCTATATCCTGAATGGTATGTT-3′ | IKe (1193–1218), AluI |
| 3′ SD mutant | 5′-cgAGATCTTCACTAGTCATGCTCATTCCTCC-3′ | IKe (1581–1553), BglII |
| 5′ EcoRI | 5′-GTATCACGAGGCCCT-3′ | pKC8 |
| 3′ KpnI | 5′-GACGAATTCCCCGGGGTA--CCAC-3′ | pKC8, EcoRI, SmaI, KpnI |
| 3′ lacZ | 5′-GGCGATTAAGTTGGGTAACGCC-3′ | pKC8 |
| 5′ 7 BglII | 5′-GAGTGGAATTCCAGATCTCCGTTTACATC-3′ | pSM7, BglII |
| 3′ 7 MstII | 5′-ACAGTATCGGCCTCAGGAAGATCGCACT-3′ | pSM7 |
| IKe toeprint | 5′-CATCGGAACACTCCAGCAG-3′ | IKe (1656–1638) |
a
Positions in oligonucleotides that differ from target sequences are indicated as follows: changes to generate restriction sites are in boldface type; point mutations are underlined; two nucleotides present in an equal mixture during synthesis are separated by a slash; nucleotides complementary to vector sequences are in lowercase letters; a deletion in the primer relative to the target is indicated by a dashed line.