TABLE 2.
Primer sequences used in this study
| Primerb | Sequencea | Purpose |
|---|---|---|
| vgrG1-Full-MluI | 5′-TCATACGCGTATGGCAGACAGCACAGG-3′ | PCR amplification of full-length VgrG1-encoding region for cloning in pBI-EGFP |
| vgrG1-Full-NheI | 5′-GGTCGCTAGCTTATAATACGGAAACCTC-3′ | |
| vgrG1-NH2-MluI | 5′-TCATACGCGTATGGCAGACAGCACAGG-3′ | PCR amplification of VgrG1 NH2-terminal-domain-encoding region for cloning in pBI-EGFP |
| vgrG1-NH2-ter-NheI | 5′-GGTCGCTAGCTTATGGGCCTCCGGCAAGCTGG-3′ | |
| vgrG1-COOH-ter-MluI | 5′-TCATACGCGTATGATGCCACCGCCGCCGCC-3′ | PCR amplification of VgrG1 COOH-terminal-domain-encoding region for cloning in pBI-EGFP |
| vgrG1-COOH-ter-NheI | 5′-GGTCGCTAGCTTATAATACGGAAACCTC-3′ | |
| vgrG1-Full-BglII | 5′-TCATAGATCTAATGGCAGACAGCACAGG-3′ | PCR amplification of full-length VgrG1-encoding region for cloning in pET-30a |
| vgrG1-Full-SalI | 5′-GGTCGTCGACTTATAATACGGAAACCTC-3′ | |
| vgrG1-NH2-ter-BglII | 5′-TCATAGATCTAATGGCAGACAGCACAGG-3′ | PCR amplification of VgrG1 NH2-terminal-domain-encoding region for cloning in pET-30a |
| vgrG1-NH2-ter-SalI | 5′-GGTCGTCGACTTATGGGCCTCCGGCAAGCTGG-3′ | |
| vgrG1-COOH-ter-BglII | 5′-TCATAGATCTAATGATGCCACCGCCGCCGCC-3′ | PCR amplification of VgrG1 COOH-terminal-domain-encoding region for cloning in pET-30a |
| vgrG1-COOH-ter-XhoI | 5′-GGTCCTCGAGTCATAATACGGAAACCTCAATC-3′ | |
| vgrG2-MluI | 5′-TCATACGCGTAATGGCAGACAGCACAGG-3′ | PCR amplification of full-length VgrG2-encoding region for cloning in pBI-EGFP |
| vgrG2-NheI | 5′-GGTCGCTAGCTCAGCCACCACCCTCCTGTCTGG-3′ | |
| vgrG2-BglII | 5′-TCATAGATCTAATGGCAGACAGCACAGG-3′ | PCR amplification of full-length VgrG2-encoding region for cloning in pET-30a |
| vgrG2-XhoI | 5′-GGTCCTCGAGTCAGCCACCACCCTCCTGTCTGG-3′ | |
| vgrG1-Full-ClaI | 5′-TCATATCGATAATGGCAGACAGCACAGG-3′ | PCR amplification of full-length vgrG1 without the stop codon for cloning in pGEN222 |
| vgrG1-Full-MluI | 5′-GGTCACGCGTTAATACGGAAACCTC-3′ | |
| vgrG1-NH2-ClaI | 5′-TCATATCGATAATGGCAGACAGCACAGG-3′ | PCR amplification of VgrG1 NH2-terminal-domain-encoding region without the stop codon for cloning in pGEN222 |
| vgrG1-NH2-MluI | 5′-GGTCACGCGTTGGGCCTCCGGCAAGCTGG-3′ | |
| bla-MluI | 5′-GGTCACGCGTATGCACCCAGAAACGCTGGTG-3′ | PCR amplification of blaM without the signal sequence for cloning in pGEN222-vgrG1s |
| bla-SalI | 5′-GGTCGTCGACTTACCAATGCTTAATCAGTG-3′ |
a
Underlined sequences represent restriction enzyme sites.
b
The designations NH2-ter and COOH-ter refer to the NH2- and COOH-terminal domains of VgrG1.