Table 1.
Primers used to generate expression constructs and PCR conditions
| Constructs | Primer names | Sequences | number of bases | cloning PCR conditions | PCR product |
|---|---|---|---|---|---|
| Construct 1 | Fwd-2-Gc | GCATCTAGAGAATTCGCCACCATGCATATTAGCCTGATGTATGCGGTGCTGTGCCTGCAGCTGTGCAGCCTGGGCTTCCTGGACAGCATCGTG | 93 |
• Initial temp: 98° C, for 30 s. 35 cycles of: • denaturation at 98° C for 10 s, • annealing at 72° C for 30 s, and • extension at 72° C for 1 min. and 30 s. |
Gc ectodomain with GPC leader, Avi-tag and His-tag |
| Rev-2-Gc | GCACTCGAGTTAGTGGTGGTGGTGGTGGTGCTCGTGCCACTCGATCTTCTGGGCCTCGAAGATATCGTTCAGGCCCACGTTGCCGAAGATGCC | 93 | |||
| Construct 2 | CCHFV-Gc-F_Ins | GCATCTAGAGAATTCGCCACCATGGCCCTGTGGATGAGACTGCTGCCTCTGCTGGCCCTGCTCGCCCTGTGGGGCCCTGACCCCGCCGCCGCTTTCCTGGACAGCATCGTG | 111 |
• Initial temp: 98° C, for 30 s. 35 cycles of: • denaturation at 98° C for 10 s, • annealing at 72° C for 30 s, and • extension at 72° C for 1 min. and 30 s. |
Gc ectodomain with human insulin leader, Avi-tag and His-tag |
| Rev-2-Gc | GCACTCGAGTTAGTGGTGGTGGTGGTGGTGCTCGTGCCACTCGATCTTCTGGGCCTCGAAGATATCGTTCAGGCCCACGTTGCCGAAGATGCC | 93 | |||
| Construct 3 | CCHFV-Gc-F_Ins | GCATCTAGAGAATTCGCCACCATGGCCCTGTGGATGAGACTGCTGCCTCTGCTGGCCCTGCTCGCCCTGTGGGGCCCTGACCCCGCCGCCGCTTTCCTGGACAGCATCGTG | 111 |
• Initial temp: 98° C, for 30 s. 35 cycles of: • denaturation at 98° C for 10 s, • annealing at 72° C for 30 s, and • extension at 72° C for 1 min. and 30 s. |
Gc ectodomain with truncated C-terminal, human insulin leader, Avi-tag and His-tag |
| CCHF-Gc-R (-51) | GCACTCGAGTTAGTGGTGGTGGTGGTGGTGCTCGTGCCACTCGATCTTCTGGGCCTCGAAGATATCGTTCAGGCCCTTCACGCTCTCCAGCCAGCAG | 97 |