Table 1.
Overview of D-fusion gene constructs cloned into the pcIpR-GOI-timm expression vector.
| D-Fusion Name (Plasmid Number) | |
|---|---|
| COOH-terminal additions: a | NH2-terminal additions: a |
| His-TAGZ-Dcoe-TEV-LL37 (p619) b | LL37-TEV-Dcoe-His (p620) |
| Dcoe-LL37 (p627) c | LL37-Dcoe (p617) c |
| Dcoe-PR39 (p625) c | PR39-Dcoe (p623) c |
| Dcoe-DEFB126ΔC (p618) d | DEFB126ΔC-Dcoe (p622) d |
| Dcoe-HBD3 (p616) | HBD3-Dcoe (p624) |
| Dcoe-HD5 (p628) | HD5-Dcoe (p621) |
| Dcoe-HNP1 (p615) [see text] | HNP1-Dcoe (p626) [see text] |
| Dcoe-YML (p674) | YML-Dcoe (p676) |
| D-YML (p675) | His-Dcoe (p614) |
| Dcoe-5EV2 (p521,p629) e | His-Dcoe- (p614*) f |
a COOH-terminal additions: the MTSK…SIV sequence for gpD and gpDcoe (see text) was followed by six amino acid spacer (see text) and then a COOH terminal addition (with the AAs arranged in their standard NH2 to COOH orientation). NH2-terminal additions: all constructs were of the arrangement M-spacer-TSK…SIV, except for p614, p614* and p676 where no spacer was included. b TEV (tobacco etch virus) represents the protease recognition sequence ENLYFQX, and the sequence ahead of gene D is recognized by the TAGZyme protease (Qiagen) for cleaving the His tag. c LL37 represents 39 AAs at the COOH-end of hCAP18 gene (167 codons) where the NH2-terminal CATHELIN domain is removed. Dcoe-LL37 fuses the sole human cathelicidin, LL37, to the COOH end of the D-spacer. Dcoe-PR39 fuses the 42 AA pig PR39 cathelicidin to a 6 AA spacer at the COOH-end of gpD. d The wild type human sperm defensin DEFB126 is expressed in the epididymis [39,40] and is 111 AAs. The molecule’s 18 COOH-terminal glycosylation sites were deleted (indicated by ΔC) in constructing Dcoe-DEFB126ΔC. The hydrophobic N-terminal end of DEFB126 is highly positively charged. e Epitopes from BVDV2 E2 protein. f As for p614, except with C to A transversion at bp 143, P48Q, in Dcoe.