TABLE 2.
Oligonucleotides used in this study
| Primer | Sequencea | Amplified region |
|---|---|---|
| oriTNF1 | 5′-ATTGGAATTCAGTTCCTCACAT-3′ | pKM101 oriT |
| oriTNR1 | 5′-CTTTTAAGCTTCATAGTACCCTCA-3′ | |
| oriTPF | 5′-CTGGAATTCAGTACACCTTGATAG-3′ | RP4 oriT |
| oriTPR | 5′-AGCAAAGCTTTTCCGCTGCATAAC-3′ | |
| kanF | 5′-GCTGCGAATCGGGAGCGGCGATAC-3′ | Part of nptII gene |
| kanR | 5′-CGCTTGGGTGGAGAGGCTATTCGG-3′ | |
| actF | 5′-TGATGGTGTTAGCCACACTGTCCC-3′ | Part of tobacco actin gene |
| actR | 5′-CTCTCAGGTGGAGCTACCACCTTA-3′ | |
| repColE1F | 5′-TGCCTCGCGCGTTTCGGTGATGAC-3′ | Part of ColE1 rep region |
| repColE1R | 5′-ACAGGTATCCGGTAAGCGGCAGGG-3′ | |
| fliCF | 5′-ACAGGATCCGCGGTAAACGACGAT-3′ | Part of E. coli fliC gene |
| fliCR | 5′-TTAAAGCTTGCCAGAAGACAGACG-3′ | |
| repANF | 5′-CAAAGCGCGTTCTCTGGTTATGTC-3′ | Part of pKM101 repA gene |
| repANR | 5′-AGCATGAGTAGTAACCCATAAGCC-3′ | |
| 27F | 5′-AGAGTTTGATCCTGGCTCAG-3′ | Universal 16S rRNA primers |
| 1492R | 5′-TACGG(T/C)TACCTTGTTACGACTT-3′ |
a
Restriction sites are in italics. F and R denote forward and reverse primers. All primers were designed in this work, apart from 27F and 1492R (16).