TABLE 1.
Primers used for domain swappinga
| Construct | Forward primer | Reverse primer |
|---|---|---|
| B110H | 5′-CCTTCCCTCCACAGGACATGCTGCGCCAGC-3′ | 5′-GTTGAACGCTGCTGCCTGGCCTGGTCTCAGGTAGAGC-3′ |
| B200H | 5′-GCCCCAAGAAGGAGAAGGAGTGCCAGCCCCCGTTTGCCTTCAGGC-3′ | 5′-GTGCAGAAGGCCTTGTCTTCACCGGGCACCCCTAACTCTCAGCAAACTTG-3′ |
| B400H | 5′-GACCAGCCCAGAGGGGACTGTGATGGCGTGCAGATC-3′ | 5′-GTGCAGAAGGCCTTGTCTTCACCGGGCACCCCTAACTCTCAGCAAACTTG-3′ |
| B600H | 5′-GGCCGATGCCGCTGCAATGTATGCGAGTGCCATTCAG-3′ | 5′-GTGCAGAAGGCCTTGTCTTCACCGGGCACCCCTAACTCTCAGCAAACTTG-3′ |
| B700H | 5′-GTGAAGGGCCCCAACATCGCCGCCATCGTC-3′ | 5′-GTGCAGAAGGCCTTGTCTTCACCGGGCACCCCTAACTCTCAGCAAACTTG-3′ |
| H110B | 5′-CCAGGGCATCCAGGGGACATGCTGGGCCTGC-3′ | 5′-TGAACGCAACTGCCTGACCTGGTCGCAGGTAAAGC-3′ |
| H200B | 5′-CCAGGGCATCCAGGGGACATGCTGGGCCTGC-3′ | 5′-CCTGAAGGCGAACGGGGGCTGGCACTCTTTC-3′ |
| H400B | 5′-CACAGGAACCAGCCCAGAGGTGACTGCGACGGCGTCCAG-3′ | 5′-CGGCCTTGTCTTCACCAAGTGCTCCTAACTCTCGGCAAACTT-3′ |
| H500B | 5′-CAGGAGCTGGAAGGAAGCTGCCGGAAGGACAACAGCTCCATC-3′ | 5′-CGGCCTTGTCTTCACCAAGTGCTCCTAACTCTCGGCAAACTT-3′ |
| H600B | 5′-AGTGGTCGTGGCCGGTGCCGCTGCAACGTGTGCCAGTGCGACC-3′ | 5′-CGGCCTTGTCTTCACCAAGTGCTCCTAACTCTCGGCAAACTT-3′ |
a
Underlines indicate human sequences. Overlapping primers were used to amplify the appropriate regions of human or bovine CD18, and the resultant products were used in the replacement amplification with either MigR1-bovine CD18 or MigR1-human CD18 as the template.