TABLE 1.
Oligonucleotide primers used for the construction of tau and ssu deletions
| Primer | Sequencea | Nucleotide positionb | Deletion(s) constructed |
|---|---|---|---|
| EE14 | 5′-GCAAGTGGAGATCTTGAACCT-3′ | 11337–11357 | tauB, tauBC |
| EE15 | 5′-ATCGGCGTAAGCTTGAGAGAT-3′ | 11895–11875 | tauB, tauBC |
| EE16 | 5′-GGTCTGCAAGCTTTACAGCGC-3′ | 13404–13424 | tauC, tauBC |
| EE17 | 5′-TCGCGCCAGCGTCGACGTTCCT-3′ | 13980–13959 | tauC, tauBC, tauABC |
| EE18 | 5′-AGAGACGTAAGATCTGGCGC-3′ | 10318–10337 | tauA, tauAB, tauABC |
| EE19 | 5′-CGGCAAGAAGCTTGTTACGCG-3′ | 10924–10904 | tauA, tauAB, tauABC |
| EE20 | 5′-TGTAGCGAAGCTTTACAGCCA-3′ | 11805–11825 | tauA |
| EE21 | 5′-AACGGTTCGTCGACTAATAAC-3′ | 12332–12312 | tauA |
| EE22 | 5′-TCGCTCCAGATCTTTGCTGGA-3′ | 250–270 | ssuA |
| EE23 | 5′-CCAGCGCAAGCTTAATGATGT-3′ | 804–784 | ssuA |
| EE24 | 5′-CATCTGGAAGCTTACTCAACTG-3′ | 1697–1718 | ssuA |
| EE25 | 5′-AGGTGAGGTCGACATCAACCTG-3′ | 2321–2300 | ssuA |
| EE26 | 5′-AAAGAGAAGATCTAACAAGTGC-3′ | 2345–2366 | ssuC, ssuCB |
| EE27 | 5′-ATAACCAAGCTTTCACTGGCG-3′ | 2916–2896 | ssuC, ssuCB |
| EE28 | 5′-ACGTGGTGAAGCTTAAACTCG-3′ | 4405–4425 | ssuB, ssuCB |
| EE29c | 5′-CTGCGTTGTCGACGAAGCAAAC-3′ | 2111–2124 | ssuB, ssuCB |
| EE30 | 5′-AGACGAGAAGCTTTCATACCG-3′ | 3693–3673 | ssuB |
| EE31 | 5′-TGGCAGCATCTGCAGATCAGC-3′ | 3063–3083 | ssuB |
| EE32 | 5′-CTGGAACCAAGCTTATCATTTG-3′ | 3641–3662 | ssuC |
| EE33 | 5′-CATCTCGAGTCGACTTAAGGC-3′ | 4189–4169 | ssuC |
| EE34 | 5′-CATGCGCGAAGCTTTTTTAAG-3′ | 12579–12599 | tauB, tauAB |
| EE35 | 5′-CGAACCTGGTCGACGCTTTTC-3′ | 13144–13124 | tauB, tauAB |
| EE36 | 5′-AAGGGCTACTGCAGTGGCGC-3′ | 12101–12120 | tauC |
| EE37 | 5′-CGGCCAGCAAGCTTTCAGCC-3′ | 12686–12667 | tauC |
| EE38 | 5′-CGCGTTAAAGCTTCGCCTGACG-3′ | 13412–13433 | tauABC |
| EE40 | 5′-AAGATACGTCGACTATGTCG-3′ | 14833–14814 | tauD |
| EE42 | 5′-ACTTAGCCCTGCAGTACGCG-3′ | 12947–12966 | tauD |
| EE43 | 5′-GGTGGCGAAAGCTTTTATCCC-3′ | 2836–2856 | ssuD |
| EE45 | 5′-TGCCCGTAAGCTTGGGTCGG-3′ | 1779–1760 | ssuD |
| EE46 | 5′-GTTGCCCTGCAGAAAGGTTCC-3′ | 1170–1190 | ssuD |
| EE47 | 5′-CGCCATCTCGAGCAACCCGC-3′ | 3362–3343 | ssuD |
| EE48 | 5′-AGCGGGGGAATTCTCAGACG-3′ | 13482–13463 | tauD |
| EE49 | 5′-GACGGAGAATTCATCGGGCG-3′ | 14247–14266 | tauD |
a
The boldface indicates changes from the template DNA for the introduction of restriction sites.
b
The position of the primers designed for tau deletions are indicated with reference to the tau operon sequence (GenBank/EBI Data Bank accession number D85613); the positions of the primers used for ssu deletions (EE29 excepted) are indicated with reference to the sequence of the ssu operon (GenBank/EBI Data Bank accession number AJ237695).
c
Primer EE29 is complementary to the E. coli pepN sequence (GenBank/EBI Data Bank accession number M15676) over the nucleotide range indicated.