TABLE 2.
Oligonucleotide primers used for cloning of chromosomal fragments and detection of gene replacement
| Primer purpose and primer | Comment | Sequencea |
|---|---|---|
| For cloning | ||
| B5 | fxuB forward primer | 5′-ACGAATTCACACGCCAGAGAACGC-3′ |
| B6 | fxuB reverse primer | 5′-CAGGATCCTGATCTGGCAATTCCAG-3′ |
| B7 | fxbA upstream reverse primer | 5′-CAGGATCCATTGGTAAGCCTTACC-3′ |
| B8 | fxbA upstream forward primer | 5′-GATCTAGAGCTGGTTCAGGGCGATG-3′ |
| U1 | fxuC forward primer | 5′-ACGAATTCGAGATCCTCGGCATCAAC-3′ |
| U5 | fxuC reverse primer | 5′-CAGGATCCTGGCGCTTCCCGAAATC-3′ |
| U6 | fxuB forward primer | 5′-GAGGATCCTGAAAGAACATATGACTC-3′ |
| U7 | fxuB reverse primer | 5′-TATCTAGACAAGATCGGCCGACATA-3′ |
| For detection of gene replacement | ||
| K1 | Out of 5′ end of aph | 5′-GAGACACAACGTGGCTTTC-3′ |
| K2 | Out of 3′ end of aph | 5′-CACGAGGCAGACCTCAGC-3′ |
| B1 | Upstream of B8 | 5′-GGTACCGACACCTCGCA-3′ |
| B9 | Out of 3′ end of fxuA | 5′-GGCGGGATCTACCTGGTAT-3′ |
| U8 | fxuC, upstream of U1 5′ end of fxb | 5′-TCCAGGCCGTGGGACGTAA-3′ |
| U9 | 5′-GTCGATGATCTGGCAATTCC-3′ |
a
Restriction sites are in bold face.