Table 3.
Primers used for gene amplification and sequencing
| Locus | Function | Putative gene product | Locus positiona | Gene size (bp) | Primersb | Primer sequence 5′-3′ | Sequence length (bp) |
|---|---|---|---|---|---|---|---|
| atpD | Energy metabolism | F0-F1 ATP synthase subunit beta | 2604717 | 1,454 | 800F | GGCCAGGACGTTCTGTTCTT | 465 |
| 1350R | CTTGAAGCCCTTGATCGTGT | ||||||
| dnaK | Stress response | Heat shock protein, 70 kDa | 126205 | 1,901 | 720F | GAAGACTTCGACATGCGTCT | 480 |
| 1400R | GCCGAGCAGCTTGTTGTC | ||||||
| glnA | Amino acid biosynthesis | Glutamine synthetase | 196923 | 1,358 | 144F | GTCATGTTCGACGGCTCCT | 474 |
| 1340R | CGCATGACTTCCTGCATCT | ||||||
| 900R | CCTTGGCATGCTTGATGAT | ||||||
| groEL | Stress response | Heat shock protein, 60 kDa | 676328 | 1,634 | 100F | GTGGTGATCAGCAGCGAAG | 504 |
| 1240R | AGGCCAAGGCCAAGAAGAT | ||||||
| 760R | CTGGAAGACATCGCCATCCT | ||||||
| rpoB polymerase | Transcription | Beta subunit RNA | 1927198 | 4,136 | 2040F | GAAAACGACGACGCCAAC | 534 |
| 3150R | TGGACCTTTTCGACCTTGTC | ||||||
| 2718R | GCGCAGAAGCTTTTCTTCC | ||||||
| trpE | Amino acid biosynthesis | Anthranilate synthase | 2262145 | 2,189 | 890F | CGCCCTATTCCTTCTTCATC | 510 |
| 2090R | ATCGATTCCGGGTGGAACT | ||||||
| 1630R | GAAATAATTCGCCAGCGTGT | ||||||
| zwf | Pentose phosphate pathway | Glucose-6-phosphate 1-dehydrogenase | 585849 | 1,475 | 530F | AGATCTTCCGCATCGACCA | 384c |
| 950R | CTTGATGGCGACGAAGGTT |
a
Gene start codon position on the circular chromosome sequence of A. tumefaciens C58 (accession number AE007869).
b
F, forward primer; R, reverse primer. Primers in boldface were used for gene sequencing in both directions. The primer denominations correspond to their hybridization regions in the gene according to the complete genome sequence of A. tumefaciens C58.
c
The size of the zwf sequences of the two strains of A. vitis was 381 bp.