TABLE 2.
Primer no. | Primer name | Sequence (5′-3′)a | Amplified fragmentb |
---|---|---|---|
1 | NIN35 | GGAGTCGGTGGTCGCAAGC | 313-bp bzdR internal fragment |
2 | REG | GTGCCAGGACTTCGAGGATGTC | |
3 | BzdN5′ | CGAATTCTGATCGAGGCCGGCATGCTG (EcoRI) | 523-bp bzdN internal fragment cloned in pK18mobbzdN |
4 | BzdN3′ | CGAATTCAGCATCTCGTTGTGCTC (EcoRI) | |
5 | BzdP5′ | CCAAGGAGCCGAGCTTCGC | 399-bp bzdP internal fragment |
6 | BzdP3′ | CCGAATTCCTTGAGCGACAGC | |
7 | δBcr5′ | GCTACGGCCGGGTGAACGTGC | 487-bp bzdQ internal fragment, bzdQ probe |
8 | δBcr3′ | CTTCACGACGCCCGGGTTCTTC | |
9 | BzdQM5′ | GCGGCGCTGTTCGGCTAC | 217-bp bzdQM intergenic region |
10 | BzdQM3′ | GATGCTGATGTAATACTCGCCGG | |
11 | EcoR8 | GGCGTCGATCCGCGCTTCC | 503-bp bzdTU intergenic region |
12 | BzdU3′ | CATCGGGAGGTAGTTGTCGCG | |
13 | EcoR11 | CTGATCGAATCCGGCTGTAC | 523-bp bzdV internal fragment |
14 | BzdV4-11 | GGAACGCGGTAGGACGGG | |
15 | BzdVW5′ | GGAACAGCTCGAACGTGGCGG | 515-bp bzdVW intergenic region |
16 | BzdVW3′ | GGGCGACGTTCATCTCCTCCATC | |
17 | BzdWXII5 | GCACCCGCAAGACGATCCG | 544-bp bzdWX intergenic region |
18 | BzdWXII3 | GCCGATCACTGTGCCGGAG | |
19 | BzdY5′ | CGAATTCAGTACAACTCCTACACCACC (EcoRI) | 366-bp bzdY internal fragment cloned in pK18mobbzdY, bzdY probe |
20 | BzdY3′ | GGAATTCTGCCGTGCTTCGGGCCGGC (EcoRI) | |
21 | YZ5′ | CGGCAAGGACGTCATCGACTTC | 322-bp bzdYZ intergenic region |
22 | YZ3′ | CCGCCGCGCCGATGCCCTG | |
23 | BzdZ-15′ | GTTGAAGGACAGGGTCGCAATCG | 447-bp bzdZ internal fragment |
24 | 8.3.4. | CTGCCCGATCGTGCCGAGC | |
25 | BzdA-25′ | GGGCACGCGCGGTGGACATC | 577-bp bzdA internal fragment |
26 | 8.3.2. | CCCCTGCCCTGTTCGAGAG | |
27 | 3REG | GGGGTACCCGTGCATCAATGATCCGGCAAG (KpnI) | 490-bp bzdRN intergenic region |
28 | N-INV-2 | GCAGTGCAGGCGATGTTGAT | |
29 | 5PBM | GGGGTACCGAAGTCCGAAGCGCTGGGTCTGC (KpnI) | 444-bp bzdQM fragment cloned in pSJ3QM |
30 | 3PBM | GCTCTAGACCCATTTTTTCCCTCCTCGGGCACTTAGTAGG (XbaI) | |
31 | 5PBZDX | GGGGTACCACATGCTCGTCGCTGCCTCCAAC (KpnI) | 602-bp bzdWX fragment cloned in pSJ3WX |
32 | 3PBZDX | GCTCTAGACCCACGCTTCGTCTCCCTTTAGCTTTCGG (XbaI) | |
33 | 5PBZDYZ | GGGGTACCGGCAAGGACGTCATCGACTTC (KpnI) | 277-bp bzdYZ fragment cloned in pSJ3YZ |
34 | 3PBZDYZ | GCTCTAGACCCATGCAAGTCCCCTTTAACCGC (XbaI) | |
35 | RIVW5′ | GCGGTACCTGCAGCAGTACGGAAAGATG (KpnI) | 313-bp bzdVW fragment cloned in pSJ3VW |
36 | RIVW3′ | GCTCTAGACCCATGGTGTCTCCCCTGGTTC (XbaI) | |
37 | 3REG | GGGGTACCCGTGCATCAATGATCCGGCAAG (KpnI) | 598-bp bzdRN fragment (PN) cloned in pSJ3PN |
38 | 5BZN | GCTCTAGACCCATCGAACTATCTCCTCTGATG (XbaI) | |
39 | 5REG | GGGGTACCGGTTTCGTCGCAGGTGCTGTCTGGC (KpnI) | 675-bp bzdR fragment (PR) cloned in pSJ3PR |
40 | 3PREG | GCTCTAGACCCATCAGCAGGTAGTTGTTCTCTTAACG (XbaI) | |
41 | AINI | CCGAATTCTGAATAGATAAGGAGAGGAGGAGCAAATGGCAGAAC (EcoRI) | 1,913-bp bzdA gene cloned in pUCBZDA |
42 | BZDB′3′ | CGCCGAACGAGTATTTCCAGCTC | |
43 | 1600R | AAGGAGGTGATCCAGCC | 1,525-bp fragment of the 16S rDNA genes |
44 | 16SF1 | GAGASTTTGATCHTGGCTCAG |
Engineered restriction sites are underlined, and the corresponding restriction enzyme is shown in parentheses.
PCR reactions were carried out with the sets of two primers indicated to the left.