Constructs used for expression of full-length, truncated, and chimeric DBL domains on surface of mammalian cells
Construct . | Description . | PCR primers* . |
|---|---|---|
| EBA175-F2(C1-C14) | AA 447-795 of EBA-175 | F: 5′ acaagtcagctggaaaagcgtgaacatattgat 3′ |
| R: 5′ tctcgtgggcccatcgtcatcacgttctttttg 3′ | ||
| EBA175-F2(C5-C14) | AA 505-795 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggcccatcgtcatcacgttctttttg 3′ | ||
| EBA175-F2(C5-C12) | AA 505-736 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggccctttataatctgaatctaattc 3′ | ||
| EBA175-F2(C5-C9) | AA 505-639 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggcccatcttcacaaggtttttcttt 3′ | ||
| R29-DBLα(C1-C14) | AA 82-391 of R29var1 | F: 5′ tctcgtcagctggagtacaccgaaggtagaaag 3′ |
| R: 5′ acgagtgggccctttccttttttgcttatcaaattgttttc 3′ | ||
| R29-DBLα(C5-C12) | AA 133-366 of R29var1 | F: 5′ tctcgtcagctggatagaaatttagaatatttgatc 3′ |
| R: 5′ acgagtgggcccacgtggacaatttaaatctataaag 3′ | ||
| FCR3-DBLγ(C1-C10) | AA 1270-1578 of FCR3varCSA | F: 5′ acttgccagctggaaaacgatggaaagaaac 3′ |
| R: 5′ acgagtgggccccctgttcaagtaatctgttg 3′ | ||
| FCR3-DBLγ(C5-C10) | AA 1315-1527 of FCR3varCSA | F: 5′ acttgccagctggtacatttcttggcaaatg 3′ |
| R: 5′ acgagtgggcccttctgtatcacacttcaattg 3′ | ||
| FCR3-DBLγ(C5-C8) | AA 1315-1517 of FCR3varCSA | F: 5′ acttgccagctggtacatttcttggcaaatg 3′ |
| R: 5′ acgagtgggccctttaacgtagtcctcgcatttc 3′ | ||
| JDP8-DBLβC2(C1-C16) | AA 853-1275 of JDP8Icvar | F: 5′ tctcgtcagctgaacaatattccaatgct 3′ |
| R: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| FCR3-DBLβC2(C1-C16) | AA 810-1260 of FCR3varCSA | F: 5′ tctcgtcagctgataaaacattctaatcgtaatctt g 3′ |
| R: 5′ acgagtgggcccttccgtatccttttcttccgc 3′ | ||
| CH1 | AA 853-1141 (DBLβ) of JDP8Icvar fused to AA 1125-1260 (C2) of FCR3varCSA | F1: 5′ tctcgtcagctgaacaatattccaatgct 3′ |
| R1: 5′ gtatttcagttccattttgctcc 3′ | ||
| F2: 5′ aaagaattacatgaacaagcac 3′ | ||
| R2: 5′ acgagtgggcccttccgtatccttttcttccgc 3′ | ||
| CH2 | AA 810-1130 (DBLβ) of FCR3varCSA fused to AA 1128-1275 (C2) of JDP8Icvar | F1: 5′ tctcgtcagctgataaaacattctaatcgtaatcttg 3′ |
| R1: 5′ gtatttatctgatattatattccattg 3′ | ||
| F2: 5′ tgtacttcgaatttggaac 3′ | ||
| R2: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| JDP8-DBLβC2(C3-C16) | AA 905-1275 of JDP8Icvar | F: 5′ tctcgtcagctgacttcgaatttggaacatttac 3′ |
| R: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| JDP8-DBLβ(C3-C7) | AA 905-1098 of JDP8Icvar | F: 5′ tctcgtcagctgacttcgaatttggaacattta 3′ |
| R: 5′ acgagtgggcccaataccacactgcgtctc 3′ |
Construct . | Description . | PCR primers* . |
|---|---|---|
| EBA175-F2(C1-C14) | AA 447-795 of EBA-175 | F: 5′ acaagtcagctggaaaagcgtgaacatattgat 3′ |
| R: 5′ tctcgtgggcccatcgtcatcacgttctttttg 3′ | ||
| EBA175-F2(C5-C14) | AA 505-795 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggcccatcgtcatcacgttctttttg 3′ | ||
| EBA175-F2(C5-C12) | AA 505-736 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggccctttataatctgaatctaattc 3′ | ||
| EBA175-F2(C5-C9) | AA 505-639 of EBA-175 | F: 5′ acaagtcagctgggaaacattgatagaatatac 3′ |
| R: 5′ tctcgtgggcccatcttcacaaggtttttcttt 3′ | ||
| R29-DBLα(C1-C14) | AA 82-391 of R29var1 | F: 5′ tctcgtcagctggagtacaccgaaggtagaaag 3′ |
| R: 5′ acgagtgggccctttccttttttgcttatcaaattgttttc 3′ | ||
| R29-DBLα(C5-C12) | AA 133-366 of R29var1 | F: 5′ tctcgtcagctggatagaaatttagaatatttgatc 3′ |
| R: 5′ acgagtgggcccacgtggacaatttaaatctataaag 3′ | ||
| FCR3-DBLγ(C1-C10) | AA 1270-1578 of FCR3varCSA | F: 5′ acttgccagctggaaaacgatggaaagaaac 3′ |
| R: 5′ acgagtgggccccctgttcaagtaatctgttg 3′ | ||
| FCR3-DBLγ(C5-C10) | AA 1315-1527 of FCR3varCSA | F: 5′ acttgccagctggtacatttcttggcaaatg 3′ |
| R: 5′ acgagtgggcccttctgtatcacacttcaattg 3′ | ||
| FCR3-DBLγ(C5-C8) | AA 1315-1517 of FCR3varCSA | F: 5′ acttgccagctggtacatttcttggcaaatg 3′ |
| R: 5′ acgagtgggccctttaacgtagtcctcgcatttc 3′ | ||
| JDP8-DBLβC2(C1-C16) | AA 853-1275 of JDP8Icvar | F: 5′ tctcgtcagctgaacaatattccaatgct 3′ |
| R: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| FCR3-DBLβC2(C1-C16) | AA 810-1260 of FCR3varCSA | F: 5′ tctcgtcagctgataaaacattctaatcgtaatctt g 3′ |
| R: 5′ acgagtgggcccttccgtatccttttcttccgc 3′ | ||
| CH1 | AA 853-1141 (DBLβ) of JDP8Icvar fused to AA 1125-1260 (C2) of FCR3varCSA | F1: 5′ tctcgtcagctgaacaatattccaatgct 3′ |
| R1: 5′ gtatttcagttccattttgctcc 3′ | ||
| F2: 5′ aaagaattacatgaacaagcac 3′ | ||
| R2: 5′ acgagtgggcccttccgtatccttttcttccgc 3′ | ||
| CH2 | AA 810-1130 (DBLβ) of FCR3varCSA fused to AA 1128-1275 (C2) of JDP8Icvar | F1: 5′ tctcgtcagctgataaaacattctaatcgtaatcttg 3′ |
| R1: 5′ gtatttatctgatattatattccattg 3′ | ||
| F2: 5′ tgtacttcgaatttggaac 3′ | ||
| R2: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| JDP8-DBLβC2(C3-C16) | AA 905-1275 of JDP8Icvar | F: 5′ tctcgtcagctgacttcgaatttggaacatttac 3′ |
| R: 5′ acgagtgggcccctcgactttgccgccatc 3′ | ||
| JDP8-DBLβ(C3-C7) | AA 905-1098 of JDP8Icvar | F: 5′ tctcgtcagctgacttcgaatttggaacattta 3′ |
| R: 5′ acgagtgggcccaataccacactgcgtctc 3′ |
AA indicates amino acid.
Two pairs of primers (F1, R1 and F2, R2) were used for PCR amplification of the 2 DNA fragments that were ligated to form the chimeric constructs CH1 and CH2.
Forward (F) and reverse (R) primers used for amplification of DNA encoding DBL domain fragments by PCR are shown