TABLE 1.
Nucleotide sequences of primers used in this worka
| Primer | Nucleotide sequence |
|---|---|
| D1 | 5′-ATGGA(a/g)TA(t/c)CA(t/c)GTAGC-3′ |
| D2 | 5′-ATGGA(a/g)TA(t/c)CA(t/c)GTTGC-3′ |
| D3 | 5′-ATGGA(a/g)TA(t/c)CA(t/c)GTCGC-3′ |
| D4 | 5′-ATGGA(a/g)TA(t/c)CA(t/c)GTGGC-3′ |
| P1 | GCCATGTGGCTAAAACTGGCC |
| P2 | CCCAAGCTTCCAAACTTTTATGTAATTATTTC |
| P3 | TGTTGCTAAAACTGGTTCAGATGAAGGGAAAGGA |
| P4 | TGATATTCCATTTTTTTATCTCCTTCTTGTCGAC |
a
The noncapitalized nucleotides represent degenerated or mutated nucleotides. The boldface nucleotides indicate differences among the RSD-PCR primers D1, D2, D3, and D4. The primers P1 and P2 were used for subcloning of the xylC gene, and the two oligonucleotides P3 and P4 were used as primers for the mutation. The underlining indicates the restriction site; the boldface indicates mutation sites.