Table I.
Sequences of the oligonucleotide primers used for the construction of recombinant Ig V genesa
| Leader-mAB13VH-D-JH gene segment | |
| H-leader (sense) | 5′ gggaagcttctcaccatgggatgg 3′ |
| H-FR4 (antisense) | 5′ gggctcgagactcaccTGAGGAGACAGTGACCA 3′ |
| H-Ov1 (antisense) | 5′ GCTGCACCTCacactggacacctgcagagaaag 3′ |
| H-Ov2 (sense) | 5′ ttctctgcaggtgtccagtgtGAGGTGCAGCTG 3′ |
| Leader-mAb13Vκ-Jκ gene segment | |
| κ-leader (sense) | 5′ gggaagcttatcaagatgaagtca 3′ |
| κ-FR4 (antisense) | 5′ gggctcgagacttacgTTTGATCTCCACCTTGG 3′ |
| κ-Ov1 (antisense) | 5′ CAACACAATTTCgcatctggaacctgcagtcagaga 3′ |
| κ-Ov2 (sense) | 5′ gcaggttccagatgcGAAATTGTGTTGACGCAGTCT 3′ |
| Recombination of germline and mutated V gene segments | |
| GIII-FR3 A (sense) | 5′ GCAATGAACAGTCTGAGAGCCG 3′ |
| GIII-FR3 B (antisense) | 5′ CGGCTCTCAGACTGTTCATTGC 3′ |
| 13-FR2 A (sense) | 5′ CCAGGGAAGGGGCTGGTGTGGGTCTC 3′ |
| 13-FR2 B (antisense) | 5′ GAGACCCACACCAGCCCCTTCCCTGG 3′ |
| κIII-FR3 A (sense) | 5′ GAAGATTTTGCAGTGTATTACTGTCAGC 3′ |
| κIII-FR3 B (antisense) | 5′ GCTGACAGTAATACACTGCAAAATCTTC 3′ |
| Mutagenized primers | |
| Thr56 A (sense) | 5′ GGGAGTACCACAAGCTACGCGG 3′ |
| Thr56 B (antisense) | 5′ CCGCGTAGCTTGTGGTACTCCC 3′ |
| Arg58 A (sense) | 5′ GGGAGTAGCACAAGGTACGCGG 3′ |
| Arg58 B (antisense) | 5′ CCGCGTACCTTGTGCTACTCCC 3′ |
a
Capital letters denote the sequences derived from the Ig V gene sequence of mAb13. Small letters denote the sequences derived from the expression vectors. Sequences of restriction sites are underlined. Codons of replacement mutations in the mutagenesis primers are indicated by underlined italic letters.