TABLE 2.
Designations and nucleotide sequences of primers used for PCR amplification and plasmid construction
| Target gene | Primera | Oligonucleotide sequence (5′ to 3′)b |
|---|---|---|
| amiE | H46(f) | CCTTATAACACTTTGATTCTTGTC |
| H58(r) | CCGCTACATAACATTGATTGGCCC | |
| H49(r) | CAAGCCCTTAGGCCCATCAACC | |
| H59(f) | CGGGATCCAGACATGGAGATATTAGTAGC | |
| H37(r) | CCAiAT(C/T)TCiGG(A/G)TA(A/G)ATiCC(A/G)TC(A/G)TC(A/G)C | |
| amiF | H69(f) | CGggatccATGGGAAGTATCGGTAGTATGGGC |
| H70(r) | GgaattcGAATCATGCCGTCATGGCAAATGCAC | |
| OI11(r) | GGAATTCATTTCCCCGGTTACAATCTCCCA | |
| H89(r) | CACGCTCACGGTATACATC | |
| 16S RNA | H276(f) | CTATGACGGGTATCCGGC |
| H676(r) | ATTCCACCTACCTCTCCCA | |
| acc(3)-IV | H121 | GGCTTTTCGCCATTCGTATTGC |
| H122 | GACGTTGGAGGGGCAAGGTCG | |
| aphA-3 | H50 | CCGGTGATATTCTCATTTTAGCC |
| H17 | TTTGACTTACTGGGGATCAAGCCTG |
a
f, forward primer; r, reverse primer.
b
Lowercase letters indicate BamHI (H69) and EcoRI (H70) restriction sites. For primer H37, two residues at the same position indicate that the oligonucleotide preparation contained a mixture of both types of molecule; “i” corresponds to inosine.