Table 1.
Intergenic junctions in chimeric LGT TP21/DEN4 and LGT E5/DEN4 constructs
| Construct | Amino acid/nucleotide sequences | LGT cDNA nt | Viability |
|---|---|---|---|
| pLGT(CpreME)/DEN4 | M A G K G L N S R | ||
| GAG CAG ATC TCT GGA AAA ATG GCC GGG AAG… …GGC TTG AAC TCG AGG | 129-2379 | No | |
| BglII XhoI | |||
| pLGT(preME)/DEN4 | L Q R R G S R R T G L N S R | ||
| 1. CTG CAGCGC CGA GGA AGT AGA AGG ACG… …GGC TTG AAC TCG AGG | 403-2379 | No | |
| PstI XhoI | |||
| N G R K R S I I D G L N S R | |||
| 2. AAC GGG AGA AAA AGA TCG ATC ATT GAC… …GGC TTG AAC TCG AGG | 426-2379 | No | |
| ClaI XhoI | |||
| NGRKRSAVDGLNSR | |||
| 3. AAC GGG AGA AAA AGG TCT GCA GTT GAC… …GGC TTG AAC TCG AGG | 428-2379 | Yes | |
| PstI XhoI | |||
| AFSLVARERGLNSR | |||
| 4. GCG TTT TCC TTG GTT GCA AGA GAG AGA… …GGC TTG AAC TCG AGG | 490-2379 | No | |
| BstBI XhoI | |||
| pLGT(NS1,2A)/DEN4 | TNSRNPTRGRRSWP | ||
| ACG AAC TCG AGG AAC CCA ACC… …AGG GGG AGA CGA TCG TGG CCT | 2382-4205 | No | |
| XhoI PvuI | |||
| pLGT(NS1,2A,2B,d3)/DEN4 | NSRNPTGTGWIRK | ||
| AAC TCG AGG AAC CCA ACC… …GGC ACG GGC TGG ATT CGA AAG | 2382-5161 | No | |
| XhoI BstBI | |||
| pLGT(NS2B,3)/DEN4 | ASRRSFNESGRRSI | ||
| GCC TCA AGA CGA TCG TTT AAT GAG… …TCT GGA AGAAGA TCT ATA | 4201-6456 | No | |
| PvuI BglII |
The terminal sequences of the corresponding cDNA fragments of LGT TP21 and LGT E5 are identical. Restriction enzyme-cleaved LGT TP21 or E5 cDNA fragments were inserted into DEN4 cDNA at appropriate sites as indicated by the underlined sequence. The amino acid and the encoding nucleotide sequences of LGT TP21 are in bold letters. (Nucleotide numbers for E5 are one less because of 5′ noncoding region deletion.) Infectivity of RNA transcripts from DNA constructs was tested by transfecting simian and mosquito cells and evaluating cell cultures for evidence of infection by immunofluorescence assay.