TABLE 1.
Summary of primers used for amplification and sequencing of MRV-EB strain cDNAsa
| Gene | Use | Primerb | Sense | Position | Sequence (5′–3′) |
|---|---|---|---|---|---|
| VP1 | RT-PCR | VP1-1 | + | 1–22 | GGC TAT TAA AGC TGT ACA ATG G |
| VP1-M1 | + | 503–523 | TTT ATA AGA GAC GGT TAG ACC | ||
| VP1-M7 | + | 1807–1827 | AAA GCG GCA AAC TCA ATT GCT | ||
| VP1-M5 | − | 915–896 | CCC TGC TTT ACT CAT AGC AA | ||
| VP1-M2 | − | 2055–2038 | TTT CGC TAT TTC GAT CCC | ||
| VP1-1-4-R | − | 3302–3282 | GGT CAC ATC TAA GCG CTC TAA | ||
| FLAC | VP1-M15 | + | 2909–2933 | CTT TAG GAG TGC CAA AAA TAG ACG C | |
| VP1-M23 | + | 3079–3098 | CCT TTT AAA GGG AAG ATA CC | ||
| VP1-M21 | − | 288–269 | GGC TAG CTT AGT TTC GAC GG | ||
| VP1-542m | − | 542–520 | GAC GCC ACG GTG ATG AAT AGG TC | ||
| Seq | VP1-M12 | + | 269–288 | CCG TCG AAA CTA AGC TAG CC | |
| VP1-M24 | + | 551–571 | ATA AGT ACG GTG TTC CAA GAC | ||
| VP1-M4 | + | 896–915 | TTG CTA TGA GTA AAG CAG GG | ||
| VP1-M18 | + | 1467–1492 | CGC AAA GTC AAC TAG AGA ATA TGC CG | ||
| VP1-M9 | + | 2349–2365 | GTC AGA TGA TGA AGT TT | ||
| VP1-M10 | + | 2955–2973 | GGT CTA TGC GCA AGA TAA A | ||
| VP1-M17 | − | 1045–1022 | CCG CAT CCA CCA TTT TCT GTT TTC | ||
| VP1-M14 | − | 1375–1354 | CCC TCC GAC CTA ACG GAA TTG G | ||
| VP1-M11 | − | 1646–1626 | TCT AAC CCC ATG ATG ATA CCC | ||
| VP1-M19 | − | 1940–1917 | GAG TTA AAT TGA AGC ACA GCG TAG | ||
| VP1-M8 | − | 2157–2138 | CAG CAC TGC AGC TTG GTC CC | ||
| VP1-2517m | − | 2517–2497 | CTG TGC TAT TCC TCG TGA CAC | ||
| VP1-M26 | − | 2943–2920 | GTA AGT GTC TGC GTC TAT TTT TGG | ||
| VP1-M25 | − | 3286–3267 | TCT AAT CTT GAA AGA AAC TGG | ||
| VP2 | RT-PCR | VP2-1 | + | 1–20 | GGC TAT TAA AGG CTC AAT GG |
| VP2-5m | + | 1337–1356 | CCA CGA ATG CAT TAC AGA AA | ||
| VP2-N2 | + | 1740–1760 | AAT TAA CTT CAG TCA CGT CCC | ||
| VP2-N1 | − | 1633–1611 | CCC TAA TCG ATT CGA AAG CAA AA | ||
| VP2-6m | − | 1991–1966 | GGT CAT CCG GAA CAC GTG CAA CAT CG | ||
| VP2-8 | − | 2681–2662 | GGT CAT ATC TCC ACA GTG GG | ||
| FLAC | VP2-N4 | + | 2440–2462 | TTT GGT AGC GAA TTA TGA CTG GG | |
| VP2-N17 | + | 2477–2499 | ACT AAG GTG TAC AAA CAA ATT CC | ||
| VP2-N16 | − | 244–221 | AGT CTT GAG AAC TTC GAG TAG CTG | ||
| VP2-N15 | − | 282–259 | AAA ATT TCA TAC TGT ACT TCT CGC | ||
| Seq | VP2-N8 | + | 465–484 | AAG ACA CAC TGC CAG ATG GT | |
| VP2-N14 | + | 800–823 | CCG CTG AAC AAC GAC ATA ATA TTC | ||
| VP2-N10 | + | 1364–1382 | CCT CAG ACT CCG TTC CAA A | ||
| VP2-N13 | + | 1785–1808 | CAG TCA TTC CAA GTC CAC AAA CGC | ||
| VP2-7m | + | 1966–1991 | CGA TGT TGC ACG TGT TCC GGA TGA CC | ||
| VP2-N11 | − | 679–656 | CCT TCT GAC CAC GCC TTC GGT CTC | ||
| VP2-N9 | − | 1165–1144 | TGC CGC AAT TAC TGT CTT AAA A | ||
| VP2-N3 | − | 2256–2234 | TTT GTC ACT TGA CCA TAG TCT CC | ||
| VP2-N18 | − | 2520–2501 | GCT CTG AAA TCA AAT TGT TG | ||
| VP3 | RT-PCR | VP3-1 | + | 1–21 | GGC TTT TAA AGC AGT ACC AGT |
| VP3-M17 | + | 281–305 | GCC AAT TTC ACC TAC AAT TTT GAG G | ||
| VP3-M5 | + | 976–992 | TTT ACC CAC GCC ATT TT | ||
| VP3-M4m | + | 1373–1397 | GTA TTT ATT CAA AAG CCA TTT AAA G | ||
| VP3-M12 | + | 2087–2111 | GAA ATA GAG AAA TAC ATT AAT ACG G | ||
| VP3-495m | − | 495–471 | GTC GCA GCG TTT TGG CAA GTG AAA G | ||
| VP3-M7 | − | 1110–1085 | CTC CAT TTT TGC CAA TCA ATG TTT CC | ||
| VP3-M2 | − | 1647–1628 | CCA AAC ATG CCC AGG CAG CC | ||
| VP3-M11 | − | 2279–2255 | GCG CTT TGA AAC TTT TAA TGA CGT C | ||
| EB-VP3-End | − | 2591–2572 | GGT CAC ATA GTG ACT GAT GT | ||
| FLAC | VP3-M10 | + | 2255–2279 | GAC GTC ATT AAA AGT TTC AAA GCG C | |
| VP3-M24 | + | 2375–2398 | AAA CTA TAC AAC GCA TTT TAC AAG | ||
| VP3-M23 | − | 307–282 | GTC CTC AAA ATT GTA GGT GAA ATT GG | ||
| VP3-495m | − | Above | Above | ||
| Seq | VP3-M1m | + | 391–416 | CAC AGA TTA TAT ATT TCC AGG CTG GG | |
| VP3-M8 | + | 1915–1942 | CAA TTA TTC GTT CGA CCT AAA GAG ATG G | ||
| VP3-M15 | − | 603–580 | CTG GCG ACT GGA AGT GCT GTA GTC | ||
| VP3-M13 | − | 1397–1373 | CTT TAA ATG GCT TTT GAA TAA ATA C | ||
| VP3-M9 | − | 1942–1915 | CCA TCT CTT TAG GTC GAA CGA ATA ATT G | ||
| VP3-2580 | − | 2580–2561 | GAC TAG TGT GTT AAG TTT TT | ||
| VP4 | RT-PCR | VP4-Fsm | + | 1–20 | GGC TAT AAA ATG GCT TCA CT |
| VP4-14 | + | 994–1019 | GGC GGT TCG CTA CCC ACT GAC TTC GG | ||
| VP4-8 | + | 1671–1690 | AGG ACT GGC CGC ATC AGT TT | ||
| VP4-9m | − | 1155–1139 | CCC TCC AGT GCA TTC GA | ||
| VP4-15 | − | 1794–1774 | CGA TGA AAC GTC TGT CCA AGC | ||
| VP4-Rsm | − | 2359–2343 | GGT CAC ATC CTC TAG AC | ||
| FLAC | VP4-7 | + | 1951–1969 | GCA CAA ATT GCA CCG AAC A | |
| VP4-22 | + | 2063–2086 | CTG ACG GAC GTT TCT TCG CAT ACC | ||
| VP4-21 | − | 217–196 | GGT ATG GTC CAT CGA GCA CTG G | ||
| VP4-6m | − | 270–251 | GGT CGG TGA GAG TAG AAT AT | ||
| Seq | VP4-16 | + | 505–528 | GCA GTG GCA AAG CAC ACA GAT CGC | |
| VP4-3 | + | 601–620 | TTA ACC GCA CAC TGC GAT TT | ||
| VP4-18 | + | 1341–1365 | GGG GCT ATA CGG TTT GCC GGC TGC G | ||
| VP4-10 | + | 1489–1513 | GAG AGA CAA CTA GGC GAA CTA CGC G | ||
| VP4-11 | − | 695–671 | GTA TTT TGG ATT GGC GGT AAT CCG G | ||
| VP4-12 | − | 1365–1341 | CGC AGC CGG CAA ACC GTA TAG CCC C | ||
| VP6 | RT-PCR | VP6-F | + | 1–20 | GGC TTT TAA ACG AAG TCT TC |
| VP6-R | − | 1356–1337 | GGT CAC ATC CTC TCA CTA TG | ||
| FLAC | VP6-1 | + | 1161–1181 | GAC AAC CTA CAA CGC GTA TTT | |
| VP6-7 | + | 1195–1216 | TTA GAA GCA TGT TGG TAA AGT G | ||
| VP6-3 | − | 268–243 | CTG GCT GAC TCA ACA TAA TTT GCG TC | ||
| VP6-6 | − | 346–323 | CCA TTT CTT TGT GAC TCT CGC ACC | ||
| Seq | VP6-4 | + | 498–521 | CCA TAT TCA GCA TCA TTC ACA CTG | |
| VP6-9 | − | 913–890 | GGT CTC ATC AAT TGA AAC GAA AGG | ||
| VP7 | RT-PCR | Beg9s* | + | 1–18 | GGC TTT AAA AGA GAG AAT |
| End9s* | − | 1062–1049 | GGT CAC ATC ATA CA | ||
| FLAC | EB-VP7-1 | + | 838–854 | GGC GGC TCA GAT GTA AT | |
| EB-VP7-4 | + | 871–891 | CCA ACA ACT GCA CCA CAA ACC | ||
| EB-VP7-3 | − | 304–281 | GAT AGT AAA GGC ATA GAG TGG AAG | ||
| EB-VP7-2 | − | 390–373 | CCC TGT TGG CCA TCC TTT | ||
| NSP1 | RT-PCR | EB-NSP1-1 | + | 1–21 | GGC TTT TTT TAT GAA AAG TCT T |
| EB-NSP1-2 | − | 1605–1581 | GGT TCA CAT TTT TTG CCG GCT AGC G | ||
| FLAC | NSP1-N3 | + | 1318–1338 | CCA TTT ACT CTC AGC TGT AAA | |
| NSP1-N9 | + | 1360–1381 | GCA TTA GTA GAG AGA TGG TAT G | ||
| NSP1-N8 | − | 223–201 | ATT GAC AGA CAT GAT GAA GTG AG | ||
| NSP1-N2 | − | 251–230 | AAA AAG CAT CTA CCA TAC TGT A | ||
| Seq | NSP1-N1 | + | 350–371 | CAA TCA ATC ATT CAG TTG TAA A | |
| NSP1-N5 | + | 716–740 | CCG TGA AGA CAC TGA TTA ACT CTG G | ||
| NSP1-N7 | + | 972–997 | CCA ACC AGC ATC TAA AGT ACG CTG CC | ||
| NSP1-N10 | − | 547–523 | GGT ACG GTA AAT TAG ATT GAT TTG C | ||
| NSP1-N6 | − | 898–874 | GAT TTA AAT TGT GCG TGA GAA ATG G | ||
| NSP1-N11 | − | 1215–1193 | AAA ACA GTG AAA TAG AAA GTG AG | ||
| NSP1-N4 | − | 1439–1420 | TCA ATT AAG CGG TTG GTT GG | ||
| NSP2 | RT-PCR | NSP2-1 | + | 1–28 | GGC TTT TAA AGC GTC TCA GTC GCC GTT T |
| NSP2-2 | − | 1056–1032 | GGT CAC ATA AGC GCT TTC TAT TCT T | ||
| FLAC | NSP2-N3 | + | 837–859 | TTT GGC AAA ATT GGC ATG CAT TT | |
| NSP2-N5 | + | 877–897 | GGG TAA TAC ATT AGA TGT GTG | ||
| NSP2-N6 | − | 270–247 | TCC GTT TCG AAA TTC ATT CCT CGG | ||
| NSP2-N2 | − | 300–281 | CAC ACA AGC ATC GCC ACC TT | ||
| Seq | NSP2-N7 | + | 427–450 | CTC TAA AGA ACT ATT GCT AAA ATC | |
| NSP2-N4 | − | 733–710 | AAC TAC ACG ATA GTG GCC CTT CCC | ||
| NSP3 | RT-PCR | NSP3-1 | + | 1–25 | GGC TTT TAA TGC TTT TCA GTT GTT G |
| NSP3-2 | − | 1072–1046 | GGT CAC ATA ACG CCC CTA TAG CCA TTT | ||
| FLAC | NSP3-N3 | + | 811–837 | GGA ACA GCA ACT GAA TTC GAT TGA TTT | |
| NSP3-N7 | + | 860–883 | GAC GAC ATT GAA ACA TTA ATT CGG | ||
| NSP3-N6 | − | 254–231 | TGT TGT TTA GAG CCT GAT CTA TGG | ||
| NSP3-N2m | − | 295–270 | GTC GCA CAT CCA ATT TCT GTT CCT AA | ||
| Seq | NSP3-N1m | + | 273–293 | GGA ACA GAA ATT GGA TGT GCG | |
| NSP3-N5 | + | 569–595 | GAG GCA TCA AAA CAG AAA ACG ACA GAG | ||
| NSP3-N4 | − | 635–612 | CAT TAG TTT TCT TAG CTT TGG CGG | ||
| NSP4 | RT-PCR | 1UG10ad† | + | 1–19 | GGC TTT TAA AAG TTC TGT TC |
| EndG10s† | − | 750–735 | GGT CAC ATT AAG ACC A | ||
| FLAC | NSP4-M5 | + | 342–365 | GAG TGG TAA AAG AAT TAA GAC AGC | |
| NSP4-M7 | + | 500–524 | AAA ACT CTA CAT GAT TGG AAA AAC G | ||
| NSP4-M3 | − | 365–342 | GCT GTC TTA ATT CTT TTA CCA CTC | ||
| NSP4-M6 | − | 450–425 | CGT ACG ATC ATC ATA TCA TAT ATT CG | ||
| Seq | NSP4-M2 | + | 480–496 | AAA CTA ATC AAA AAG CG | |
| NSP4-M1 | − | 288–274 | CCA AGC CTC AGC AAA | ||
| NSP5 | RT-PCR | NSP5-5m | + | 1–20 | GGC TTT AAA AGC GCT ACA GT |
| NSP5-3 | − | 664–646 | GGT CAC AAA ACG GGA GTG G | ||
| FLAC | NSP5-M1 | + | 280–303 | GCT GGC GTG TCT ATG GAT TCA TGC | |
| NSP5-M6 | + | 396–420 | GGA CAC CAC AAG GTC AAA AAT TGC G | ||
| NSP5-M5 | − | 239–217 | CTG GTG AGT GGA TCG TTC GAA GC | ||
| NSP5-M2 | − | 354–331 | GGC GAG ATC CAC TTG ATT GCA TCC |