Article Figures & Data
Figures
- FIG 1
Schematic map of position coordinates for all sequenced loci in EEHV3, EEHV4, EEHV5A, EEHV5B, and EEHV6 compared to the complete EEHV1A(Kimba) genome. The diagram is drawn to scale with bars representing all of the DNA sequence blocks generated here aligned relative to the complete 177,316-kb genome of EEHV1A(Kimba) (13). The data reported by Ehlers et al. (11) for EEHV1B(Kiba) is also given for comparison in the top line. The locations of the predicted Ori-Lyt dyad symmetry locus (black circle), the large 40-kb inverted (Inv) core domain III-II-I segment (green arrow), the putative immediate early-like ORF-L transactivator protein coding region (blue arrow), and the three major hypervariable domains CD-I, CD-II, and CD-III (yellow boxes) as described by Richman et al. (10) are indicated.
- FIG 2
Radial phylogenetic tree showing evolutionary relationships between EEHV1, EEHV2, EEHV3, EEHV4, EEHV5, and EEHV6 in the highly conserved U38(POL) DNA gene. The diagram shows a distance-based Bayesian nearest-neighbor evolutionary tree dendrogram produced in MEGA5 illustrating the branching patterns of the GC-rich (EEHV3 plus EEHV4) compared to the AT-rich EEHV groups and of the two distinct subgroups (EEHV1 plus EEHV6 compared to EEHV2 and EEHV5) within the AT-rich branch. After several small gaps and nonaligned nucleotides were omitted, the final data set for the segment of U38(POL) gene DNA used that is common to all eight EEHV types was 1,077 bp. The number of nucleotide substitutions per site is shown by the bar.
- FIG 3
Evaluation of EEHV1A-1B chimeric domain CD-I patterns and boundaries relative to EEHV6. The diagrams show SimPlot comparisons of the nucleotide identity patterns between EEHV6, EEHV1A, EEHV1B, and EEHV2 across the 3.0-kb EEHV1B chimeric domain CD-I. (a) CD-I. The 5,000-bp U39(gB)-U38(POL) segment from EEHV1A(Kala, NAP18) map coordinates 73,959 to 79,043 compared to EEHV6(NAP35) (blue) and to EEHV2(Kijana, NAP12) (gray) is shown. (b) CD-I. The 4,800-bp U39(gB)-U38(POL) segment from EEHV1A(Kala, NAP18) map coordinates 73,987 to 78,860 compared to EEHV1B(Kiba, NAP14) (red) and to EEHV6(NAP35) (blue) is shown. (c) CD-I. The 5,000-bp U39(gB)-U38(POL) segment from EEHV1A(Kala, NAP18) map coordinates 73,959 to 79,043 compared to EEHV1B(Haji, NAP19) (green) and to EEHV6(NAP35) (blue) is shown. Arrows mark the positions of the chimeric domain boundary transitions, and the relevant DNA accession numbers are included in Table S1 in the supplemental material.
- FIG 4
Evaluation of EEHV1A-1B chimeric domain CD-II and CD-III patterns and boundaries relative to EEHV6. The diagrams show SimPlot comparisons of the nucleotide similarity patterns between EEHV6, EEHV1A, and EEHV1B across both the left-hand side (LHS) and right-hand side (RHS) boundaries of the 3.7-kb CD-II region and across the LHS boundary only of the 8.3-kb CD-III region of EEHV1B. (a) CD-II LHS. The left-hand side of the 2.6-kb U27(PPF)-U47(gO) segment across map coordinates 101,128 to 103,703 for EEHV1A(Kala, NAP18) compared to EEHV1B(Kiba, NAP14) (red) and to EEHV6(NAP35) (blue). (b) CD-II RHS. The right-hand side of the 2.7-kb U48(gH)-U50(PAC2) segment across map coordinates 105,364 to 108,084 for EEHV1A(Kala, NAP18) compared to EEHV1B(Kiba, NAP14) (red) and to EEHV6(NAP35) (blue) is shown. (c) CD-III LHS. The left-hand side of the 4.3-kb U77-U82.5(ORF-O) segment across map coordinates 143,644 to 147,936 for EEHV1A(Kala, NAP18) compared to EEHV1B(Kiba, NAP14) (red) and to EEHV6(NAP35) (blue) is shown. (Data for the right-hand segment of CD-III are not yet available for EEHV6.) Arrows mark the positions of the chimeric domain boundary transitions, and the relevant DNA accession numbers are included in Table S1 in the supplemental material.
- FIG 5
Evaluation of the EEHV5A-5B chimeric domain patterns and boundaries relative to EEHV2. The diagrams show SimPlot comparisons between EEHV5A, EEHV5B, and EEHV2 across all three EEHV5B chimeric domain regions, including all of CD-I (2.4 kb) and CD-II (1.1 kb) and the left-hand side boundary of CD-III (>2.7 kb). (a) CD-I. The 4,800-bp U39(gB)-U38(POL) segment from across map coordinates 74,002 to 78,861 for EEHV5A(NAP50) compared to EEHV5B(NAP58) (blue) and to EEHV2(NAP12) (red) is shown. (b) CD-II. The 2.8-kb U48(gH)–U48.5(TK)–U49–U50 segment from across map coordinates 105,396 to 108,178 for EEHV5A(NAP50) compared to EEHV5B(NAP58) (blue) and to EEHV2(NAP12) (red) is shown. (c) CD-III. The left-hand side only of the 4,300-bp segment across map coordinates 143,644 to 147,958 for EEHV5A(NAP50) encompassing U77(HEL C terminus), U77.5(ORF-M), U80.5(ORF-N), U81(UDG), U82(gL), and U82.5(ORF-Oex3) compared to EEHV5B(NAP58) (blue) and EEHV2(NAP12) (red) is shown.
- FIG 6
Linear DNA level phylogenetic trees comparing four gene loci from EEHV3, EEHV4, EEHV5A, EEHV5B, and EEHV6 with their orthologues in EEHV1, EEHV2, and other key herpesviruses. The diagrams present linear Bayesian maximum likelihood phylogenetic dendrograms at the nucleotide level for a set of representative EEHV gene loci from the five EEHV3, EEHV4, EEHV5A, EEHV5B, and EEHV6 (proposed Deltaherpesvirinae [δ]) genomes determined here, together with orthologues from EEHV1A, EEHV1B, and EEHV2 (10). These are compared with matching segments of selected orthologous gene loci among representative herpesviruses from all three other mammalian subfamilies, Alphaherpesvirinae (α), Gammaherpesvirinae (γ), and Betaherpesvirinae (β). Alternative virus names and GenBank accession numbers for both the non-EEHV and EEHV genome file sources used are listed in supplemental material or in Table S1 in the supplemental material. Bootstrap values are shown as percentages at the nodes. The final DNA segment sizes were as follows for panels a to d: (a) U38(POL) locus, DNA polymerase, 927 bp; (b) U73(OBP) locus, origin binding protein, 576 bp; (c) U76(POR)-U77(HEL) locus, portal protein plus helicase subunit; (d) U71-U72(gM) locus, myristylated tegument protein plus glycoprotein M, 353 bp. All four panels use Marek's disease alphaherpesvirus (MDV) as an outgroup.
- FIG 7
Linear protein level phylogenetic trees comparing five protein segments from EEHV5 and EEHV6 with their orthologues in EEHV1, EEHV2, and other key herpesviruses. The diagrams present linear Bayesian maximum likelihood phylogenetic dendrograms at the amino acid level for a set of representative proteins from the prototype EEHV5A, EEHV5B, and EEHV6 genomes, as well as up to five other orthologues from EEHV1A, EEHV1B, and EEHV2 (proposed Deltaherpesvirinae [δ]). These are compared to matching segments of orthologous gene loci from selected herpesviruses representative of the Alphaherpesvirinae (α), Gammaherpesvirinae (γ), or Betaherpesvirinae (β) subfamilies. Bootstrap values are shown as percentages. The final protein segment sizes used were as follows for panels a to e: (a) U48.5(TK), thymidine kinase, 244 aa, with turtle herpesvirus (TurtleHV) used as the outgroup; (b) U27(PPF), polymerase processivity factor, 110 aa, with MDV used as the outgroup; (c) U39(gB), glycoprotein B, 727 aa, including only the Betaherpesvirinae as references with EBV used as the outgroup; (d) U82(gL), glycoprotein L, 85 aa, with only Betaherpesvirinae included as references and EBV used as the outgroup; (e) U81(UDG), uracil DNA glycosylase, 252 aa, with only Roseolovirus species used for reference and HHV7 used as the outgroup.
- FIG 8
Comparison of EEHV POL protein divergence levels with great ape plus New World and Old World primate versions within the alpha-, beta-, and gammaherpesvirus subfamilies. The diagram shows a linear distance-based Bayesian phylogenetic tree dendrogram at the amino acid level for each of the eight prototype EEHV genomes (proposed Deltaherpesvirinae [δ]) across the largest segment of U38(POL) in common (1,080 bp, Kimba equivalent coordinates 77,783 to 78,863). To allow direct comparisons of estimated ages of divergence (see the text), these are compared with matching segments of orthologous gene loci from selected primate herpesviruses representative of just the Simplexvirus (Alphaherpesvirinae [α]), Lymphocryptovirus genus (Gammaherpesvirinae [γ]), and Cytomegalovirus (Betaherpesvirinae [β]) genera. Human VZV (Varicellovirus genus) was used as the outgroup. The evolutionary history was inferred by using the maximum likelihood method based on the Kimura two-parameter model with initial trees for the heuristic model obtained by applying the neighbor-joining method. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. Evolutionary analyses were conducted in MEGA5. Bootstrap values are shown as percentages.
- FIG 9
Clustal alignment illustrating variability patterns among the intact glycoprotein L (gL) and uracil DNA glycosylase (UDG) proteins from distinct types of EEHV genomes. Clustal-W presentations comparing the predicted primary amino acid sequence alignments of the prototypes of all six available types and subtypes of AT-rich branch Proboscivirus genomes compared to their closest orthologues in either all Betaherpesvirinae or in just the Roseolovirus genus. GenBank accession numbers for the EEHV proteins and betaherpesvirus reference species used are given in the supplemental material. (a) U82(gL), glycoprotein L, intact proteins; (b) U81(UDG), uracil DNA glycosylase, intact proteins. Positions with 100% similarity are boxed. Gaps introduced to maximize alignment are indicated by hyphens.
- FIG 10
Clustal alignments illustrating variability patterns among the intact ORF-J, glycoprotein N (gN), and ORF-N(vCXCL1) proteins of AT-rich branch EEHV genomes. Clustal-W presentations comparing the primary amino acid sequence alignments for all available predicted Proboscivirus versions of three small unique or poorly conserved proteins that do not have orthologues in other herpesviruses for comparison are shown. (a) U45.7(ORF-J), intact novel proteins. (Data are not yet available for ORF-J of EEHV5A or EEHV5B.) (b) U46(gN), glycoprotein N, intact proteins. (Data are not yet available for gN of EEHV5A or EEHV5B.) (c) U80.5(vCXCL1, ORF-N), possible inhibitory alpha-chemokine ligand. [The vCXCL1(ORF-N) gene is absent from EEHV1B.] Positions with 100% similarity are boxed.
Tables
- TABLE 1
Summary of six EEHV-positive elephant cases evaluated in these studies
Case Virus type Strain Elephant namea Host species, sex, and ageb Location Yr Pathologyc (reference) DNA sourced Sequenced DNA (bp) 1 EEHV3 NAP27 Hansa EM, F, 7y Seattle, WA 2007 Fatality (3) Necropsy tissue sample 4,117 2 EEHV4 NAP22 NA EM, F, 5y Oklahoma 2004 Fatality (3) Necropsy tissue sample 5,743 3 EEHV5A NAP28 NA EM, F, 69y Washington, DC 2007 Routine (4) WB sample 14,615 4 EEHV5A NAP50 Methai2 EM, F, 40y Texas 2011 Letharg. (17) WB sample 26,090 5 EEHV5B NAP58 Tucker3 EM, M, 8y Texas 2011 Asympt. (17) WB, TW samples 29,347 6 EEHV6e NAP35 NA LA, F, 15m Arkansas 2009 Sympt. (4) WB sample 31,828 Total 111,749 ↵a NA, not available.
↵b The host animal species (Elephas maximus [EM] or Loxodonta africana [LA]), sex (female [F] or male [M]), and age (in months [m] or years [y]) is shown.
↵c Letharg., lethargic; Asympt, asymptomatic; Sympt, symptomatic.
↵d WB, whole blood; TW, trunk wash fluid.
↵e Survived after FCV treatment.
- TABLE 2
Summary of PCR-sequenced EEHV3, EEHV4, EEHV5A, EEHV5B, and EEHV6 gene coding regions
Gene/ORF no./IDa HCMV ORF HSV ORF Orientationb Protein name Statusc Presence/absence of the indicated ORFd EEHV3 NAP27 EEHV4 NAP22 EEHV5A NAP28 EEHV5A NAP50 EEHV5B NAP58 EEHV6 NAP35 U39 UL55 UL29 F gB Core + + + + + U38 UL54 UL30 F POL Core + + + + + + U33 UL49 Nil F Cys-rich β/γ + + + + U28 UL45 UL39 F RRA Core + + + + U27.5/ORF-H Nil UL40 F RRB α/β2 + + + + U27/ORF-I UL44 UL42 F PPF Core + + + U45.7/ORF-J Nil Nil F Novel + U46 UL73 UL49A F gN Core + U47/ORF-D UL74 Nil R gO β + U48 UL75 UL22 R gH Core + + + + U48.5/ORF-E Nil UL23 R TK α/γ + + + + U49 UL76 UL24 F Core + + + + U50 UL77 UL25 F PAC2 Core + + + U51 UL78 Nil F vGPCR1 β + + + + U57 UL86 UL19 R MCP Core + + + + U60ex3 UL89 UL15ex2 R TERex3 Core + + + + + U62 UL91 Nil F β/γ + + + U63 UL92 Nil F β/γ + U66ex2 Nil Nil R TERex2 Core + U66ex1 UL89ex1 UL15ex1 R TERex1 Core + U70 UL98 UL12 F EXO Core + + + + + + U71 UL99 UL11 F myrTeg. Core + + + + + + U72 UL100 UL10 R gM Core + + + + + + U73/ORF-G Nil UL09 F OBP α/β + + + + + + U76 UL104 UL06 R POR Core + + + + + + U77 UL105 UL05 F HEL Core + + + + + + U77.5/ORF-M Nil Nil F Nuclear Novel + + + U80.5/ORF-N Nil Nil R vCXCL1 Novel + + + U81 UL114 UL02 R UDG Core + + + U82 UL115 UL01 R gL Core + + + U82.5/ORF-Oex3 Nil Nil R S/TGlyP Novel + + + U82.5/ORF-Oex2 Nil Nil R S/TGlyP Novel + + + U82.5/ORF-Oex1 Nil Nil R S/TGlyP Novel + + + U83.5/ORF-Pex2 Nil Nil R S/TGlyP Novel + U83.5/ORF-Pex1 Nil Nil R S/TGlyP Novel + U84.5/ORF-Q Nil Nil R S/TGlyP Novel A U85.5/ORF-Kex3 Nil Nil R SplGlyP Novel + + + U85.5/ORF-Kex2 Nil Nil R SplGlyP Novel + + + U85.5/ORF-Kex1 Nil Nil R SplGlyP Novel + + + U86.5/ORF-L Nil Nil R IE-like Novel + + + ↵a ID, identification.
↵b F, forward; R, reverse.
↵c Novel, not found in any other herpesviruses; β, betaherpesvirus subfamily only; Core, common to all herpesvirus subfamilies; β/γ, betaherpesvirus and gammaherpesvirus subfamilies only; α/β, alphaherpesvirus and betaherpesvirus subfamilies only; α/β2, alphaherpesvirus subfamily and roseoloviruses only.
↵d +, partial or intact ORF present (see Table S1 in supplemental material for detailed coordinates, percent divergence, and GenBank accession numbers); A, gene absent.
- TABLE 3
DNA and protein divergence in ORFs across EEHV3, EEHV4, and EEHV1A
Gene locus EEHV1A coordinatesa Protein size (no. of aa)b Nucleotide level divergence (%)c Amino acid level divergence (%)c Overall % G+C contentd Wobble % G+C contentd 3/4 3/1A 4/1A 3/4 3/1A 4/1A 3 4 1A 3 4 1A U38(POL) (77782–78912) (412) 7.0 34 34 7.5 26 25 67 68 42 89 89 43 U60(TERex3) (123721–124037) (105) 3.8 22 23 1 5 5 57 56 42 91 86 41 U71(MyrTeg) 132954–133241 96 26 50 49 29 63 66 61 58 46 76 61 41 U72(gM) (133320–133608) (95) 14 45 13 52 55 40 80 42 U72(gM) (133316–134404) 372 34 35 55 40 89 49 U73(OBP) (134645–135283) (237) 17 48 22 39 63 41 91 46 U73(OBP) (134404–135284) (438) 54 54 64 44 88 49 U76(POR) (140967–141295) (104) 3.3 34 30 1 21 21 64 63 42 98 99 47 U77(HEL) (141246–141878) (189) 3.5 30 30 2.5 23 22 67 67 41 96 97 50 ↵a The EEHV1A coordinates are based on Ling et al. (13). Entries shown in parentheses indicate that an incomplete ORF or smaller region was used.
↵b aa, amino acids.
↵c Divergence of EEHV3, EEHV4, and EEHV1A is indicated as follows: EEHV3 and EEHV4 (3/4), EEHV3 and EEHV1A (3/1A), and EEHV4 andEEHV1A (4/1A).
↵d The total guanine-plus-cytosine nucleotide percentages of EEHV3, EEHV4, and EEHV1A (indicated by 3, 4, and 1A in the table) compared to those at just the third nucleotide position in all codons from the predicted proteins.
- TABLE 4
DNA and protein divergence in ORFs across the EEHV1A, EEHV1B, and EEHV6 genomesa
Gene locus EEHV1A(Kimba) coordinatesb Protein sizeb Nucleotide level divergence (%)c Amino acid level divergence (%)c Chimeric domain 1A-1B 1A-6 1B-6 1A-1B 1A-6 1B-6 U39(gB) 73959–76511 836 21 16 20 14 11 13 CD-I U38/(POL) (76544–79043) (832) 5 17 16 4 12 12 CD-I U33 (83628–84428) (266) 1.3 22 21 0.3 19 18 U28(RRA) 99358–99760 801 1.1 14 15 0 8 8 U27.5(RRB) 99804–100709 302 1.4 14 14 0.3 17 17 U27(PPF) 100960–102186 408 1.8 16 16 0.5 6 6 U45.7(ORF-J) 102168–102775 168 26 16 25 32 14 33 CD-II U46(gN) 102759–103713 96 29 16 28 20 6.2 18 CD-II U47(gO) 103075–103713 212 35 16 35 38 11 37 CD-II U48(gH) (105363–105903) (179) 31 17 28 33 21 37 CD-II U48.5(TK) 105835–106908 356 13 16 18 16 21 20 CD-II U49 106910–107608 232 3.6 15 15 0.4 12 13 U50(PAC2) (107427–108083) (220) 4.0 23 23 3.1 20 21 U51(vGPCR1) (109398–110239) (286) 4.2 22 23 3.3 14 17 U57(MCP) (115577–117866) (748) 2.7 17 17 1.2 7 7 U60(TERex3) 123601–124175 (194) 2.9 12 13 0d 0d 0d U62 124231–124477 88 0.8 13 13 0 9 9 U71(MyrTeg) 132954–133241 96 8 25 25 9 23 26 U72(gM) (133320–133614) (97) 4.4 13 14 3.3 9.2 7.2 U73(OBP) (134615–135415) (277) 3.0 14 14 1.9 6.0 6.7 U76(POR) (139998–141295) (433) 1.9 13 13 0 4.2 4.2 U77(HEL) (141294–141864) (190) 2.8 12 13 0.5 7.0 7.5 U77.5(ORF-M) 143988–145502 503 16 16 19 13 13 16 CD-III U80.5(ORF-N) 145642–145959 106 Del 31 Del Del 29 Del CD-III U81(UDG) 146027–146980 317 29 19 30 29 19 30 CD-III U82(gL) 146946–147860 304 31 18 30 33 22 41 CD-III U82.5(ORF-O) (147700–147910) (78) 33 16 31 34 18 37 CD-III U85.5(ORF-K) (152200–153350) (380) 1.7 26 26 0.8 25 25 U86.5(ORF-L) (155324–158615) (1084) 2.4 11 11 0.6 18 18 ↵a The rows in the table shown in bold type indicate ORFs that are highly variable between EEHV1A and EEHV1B.
↵b The EEHV1A coordinates are based on Ling et al. (13). The values shown in parentheses indicate that an incomplete ORF or shorter region was used.
↵c The divergence between EEHV1A and EEHV1B (1A-1B), EEHV1A and EEHV6 (1A-6), and EEHV1B and EEHV6 (1B-6) is shown. Del, ORF-N is absent (deleted) in all EEHV1B strains.
↵d All three TER proteins are identical or only 1 aa different over this 194-aa segment.
- TABLE 5
DNA and protein divergence in ORFs across EEHV5A, EEHV5B, EEHV2, and EEHV1Aa
Gene locus EEHV1A coordinatesb Protein sizeb Nucleotide level divergence (%)c Amino acid level divergence (%)c Chimeric domain 5A-5B 5A-2 5B-2 5A-1 5A-5B 5A-2 5B-2 5A-1 U39(gB) (74091–76511) (837) 10 16 15 27 4.2 7.6 6.3 21 CD-I U38(POL) (76583–79038) (818) 1.3 18 17 24 1.4 11 11 21 CD-1 U33 (83648–84183) (176) 0.8 28 28 37 0 25 25 43 U28(RRA) (98637–99760) (374) 0.4 17 17 23 0 8.6 8.6 16 U27.5(RRB) 99804–100709 (301) 0.9 16 16 24 0.7 6.3 6.3 15 U27(PPF) (100960–101609) (216) 0.8 19 19 31 0.9 14 14 25 U48(gH) (105396–105911) (172) 0 18 18 38 0 16 16 34 U48.5(TK) 105835–106908 344 14 18 20 30 13 19 19 27 CD-II U49 106910–107628 233 6.0 11 10 24 2.1 6.9 4.7 24 CD-II U50(PAC2) (107428–108178) (261) 0.2 18 18 32 0 19 19 34 U51(vGPCR1) (109225–110065) (292) 4.6 22 23 32 2.7 17 17 25 U57(MCP) (116671–117809) (380) 0.2 19 19 27 0.3 9.2 9.2 19 U60(TERex3) (123674–124180) (169) 1.2 13 13 21 0 1.3 1.3 3.0 U62 (124231–124434) (75) 0.5 16 16 29 0 10 10 19 U71(MyrTeg) 132954–133241 98 8.7 24 23 44 14 27 30 56 U72(gM) 133316–133613 (101) 0.3 16 17 21 0 10 10 12 U73(OBP) (134605–134430) (269) 0 12 12 21 0 4.4 4.4 10 U76(POR) (139815–141295) (493) 0.2 13 13 22 0.4 4.1 4.1 7.8 U77(HEL) (141294–141793) (181) 0.4 14 14 24 0.7 6.0 6.0 14 U77.5(ORF-M) 143988–145511 484 1.1 16 17 29 0 10 10 17 CD-III U80.5(ORF-N) 145642–145959 106 28 23 27 38 32 24 33 47 CD-III U81(UDG) 146027–146980 324 27 23 28 33 23 21 20 29 CD-III U82(gL) 146946–147776 312 27 17 27 34 26 16 26 44 CD-III U82.5(ORF-Oex3) 147694–147958 (88) 23 21 25 32 21 20 20 25 CD-III U82.5(ORF-O) 147700–148985 487 36 54d 34 53d U83.5(ORF-P) 146932–150638 526 33 48d 33 52d U85.5(ORF-K) 152042–154436 (678) 1.3 23 22 41 0.4 29 27 48 U86.5(ORF-L) 155053–155356 (97) 1.4 10 12 30 1.0 7.0 8.4 20 ↵a The prototype genomes used were EEHV5A(NAP50), EEHV5B(NAP58), EEHV2(NAP12), and EEHV1A(NAP18). The rows in the table shown in bold type indicate ORFs with high-level variability (hypervariable genes) between EEHV5A and EEHV5B.
↵b The values in parentheses indicate that incomplete ORFs were used.
↵c The divergence between EEHV5A and EEHV5B (5A-5B), EEHV5A and EEHV2 (5A-2), EEHV5B and EEHV2 (5B-2), and EEHV5A and EEHV1 (5A-1) is shown.
↵d These values show divergence between EEHV5B and EEHV1A.
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
Section S1 (Detailed ORF coordinate data for individual EEHV PCR loci.)
Table S1 (Details of sequenced EEHV3, EEHV4, EEHV5A, EEHV5B, and EEHV6 gene coding regions.)
Section S2 (Examples of primers used to amplify specific gene loci.)
Section S3 (Full or alternative virus names and non-EEHV GenBank accession numbers.)
Section S4 (EEHV GenBank accession numbers for EEHV DNA or protein files used in the phylogenetic dendrograms or Clustal comparisons.)
PDF, 206K
- Supplemental file 1 -
