Table 1.
PCR primers used in this study
| Primer | Sequencea | Position (nt) in AdDNV | Target or use |
|---|---|---|---|
| AdVPR | TTTGTTGCAATCCCATAATAGTAC | 2610–2633 | Near 3′ end of VP mRNA |
| NAdR | gctctagatCATCTTGAACGTTTACCACCACT | 3892–3915 | Just upstream of VP4 |
| Adsp | tcggaattcCACGTTCTTGTGGATGAGG | 4362–4380 | 19-37 nt into VP2 |
| Adsvp | gccTACCAGAAATCCGTGTAATGACA | 4546-4534/4403-4393 | Small intron splice |
| Ad3s | CGTGAGTACTGATACTTTTTATTT | 4435–4412 | End of ORF-B (TGA) |
| Ads | gccCCTCAACAACCTAAAAAACGTGAGTACTGA | 4453–4424 | End of ORF-B (TGA) |
| Ad6s | CCTAAAAAACGTGAGTACTGA | 4444–4424 | End of ORF-B (TGA) |
| Adl | gccGACGTAATTGGTGGACCTGTATATCCT | 4477–4451 | End of ORF-B (−8 aa) |
| Adm | gccCCTGTATATCCTCAACAACCTAAA | 4462–4439 | End of ORF-B (−4 aa) |
| AdlgF | tcggaattcATGTCTGGCGTCTTTACA | 5230–5213 | Start of VP1 (ORF-B) |
| RNAtag | gggtctagagctcgagT17 | Poly(A) sequence | 3′/5′-RACE (first round) |
| ADAP | gggtctagagctcgagT | Subsequent rounds of RACE | |
| ProNSF | acggtaccGATATAAAGAGCAAGCACCC | 109–128 | NS1 promoter (KpnI) |
| ProNSR | gaagatCTGTGCTGGAGGCGCTTCTACTGCAGCGAACAACGTACCTGAGTTCCAGAACAC | 240–207 | NS1 promoter (reverse) |
| ProVP1R | gaagatctAAGACGCCAGAGATTTAATACT | 5217–5238 | VP1 (BglII site; reverse) |
| ProVP1F | gtaggtaccGATATAAAGAGCAAGCACCCA | 5323–5300 | VP1(KpnI site; forward) |
| PrNSMf | ACGGTACCGACTATAAATAGAGCTGAGC | MlDNV (forward) | |
| PrNSMr | GAAGATCTATCTTGCAATAGATATACCTA | MlDNV (reverse) | |
| pFECRIF | CGCAAATAAATAAGAATTCTACTGTTTTCGTAAC | Mutation in pFastBac1 | |
| pFECRIR | GTTACGAAAACAGTAGAATTCTTATTTATTTGCG | Mutation in pFastBac1 | |
| Ad1HAR | GCTCTAGATCAAGCGTAATCTGGAACATCGTATGGGTATTTTTGTTGCAATCCCATAATA | VP products (reverse primer [HA]) | |
| AdATG1B | ggaattcATGTCCGTCTTTACAGATCTCAC | ||
| AdmATG1B | ggaattcACCTCCGTCTTTACAGATCTCTCAC |
a
The AdDNV sequence is shown in capital letters. Underlined nucleotides indicate stop codons (e.g., TGA) or restriction sites (e.g., ggtacc [KpnI]).