TABLE 1.
Oligonucleotides used in PCR
| Primer | Sequencea | Positionb (nt) |
|---|---|---|
| ssg1 | 5′ GGCGAATTCGAACAGCTACGTGGCGAAGTCGCCA 3′ EcoRI | −194 to −170 |
| ssg2 | 5′ GTGGGATCCGTGCTCGCGGCGCTGGTCGTCTC 3′ BamHI | +539 to +517 |
| ssg3 | 5′ GGGAATTCCATATGCGCGAGTCGGTTCAAGCA 3′ EcoRI NdeI | −30 to −10 |
| ssgN3 | 5′ GGGAATTCCATATGATGACGTTCCTCGTCTCCG 3′ EcoRI NdeI | +1 to +22 |
| ssg4 | 5′ GTGGATCCCCGGTCAGCCGGCGTTCTG 3′ BamHI | +412 to +394 |
| Q1 | 5′ CTGAATTCTAGCATCGAGGGCAGGACATCA 3′ EcoRI | −1450 to −1427 |
| Q6 | 5′ CTGAAGCTTAACGACCGGCCCAGGTGGCGAC 3′ HindIII | +520 to +541 |
| Q10 | 5′ CTGAATTCGGCTCAGCGGTTGCAGAACGAG 3′ EcoRI | −196 to −175 |
| Q11 | 5′ ATAGGGATCCCGGGTCTCGTAGCGCAGC 3′ BamHI | +75 to +48 |
| Q14 | 5′ CTGGATCCTGGTGCCGCTGGGGCAGGAGG 3′ BamHI | +310 to +330 |
| Q15 | 5′ CTGAAGCTTGAGACGGGTTACGGCCACGATGC 3′ HindIII | +1870 to +1850 |
a
Underlined nucleotides indicate nonhomologous sequences added to create restriction sites (in italics) at the ends of the PCR fragments.
b
Location with respect to the first ucleotide (+1) of the ATG translational start codon of S. griseus ssgA of S. coelicolor ssgA for oligonucleotides designated “ssg” or “Q”, respectively.