JOURNAL OF VIROLOGY, Mar. 1982, p. 931-939 Vol. 41, No. 30022-538X/8V030931-09$02.0O/0

DNA Of Herpesvirus Pan, a Third Member of the Epstein-Barr Virus-Herpesvirus Papio GroupMARK HELLER,1 PAUL GERBER,2t AND ELLIOTT KIEFF1*

Marjorie B. Kovler Laboratories, The University of Chicago, Chicago, Illinois 60637,1 and Division ofBiological Standards, National Institutes of Health, Bethesda, Maryland 202052

Received 13 August 1981/Accepted 23 October 1981

The DNA of herpesvirus pan, a primate B-lymphotropic herpesvirus, sharesabout 40%o well-conserved sequence relatedness with Epstein-Barr virus (EBV)and herpesvirus papio DNAs. Labeled cloned fragments from the EBV recombi-nant DNA library were cross hybridized to blots of EcoRI, XbaI, and BamHIrestriction endonuclease fragments of herpesvirus pan DNA to identify and maphomologous sequences in the herpesvirus pan genome. Regions of colinearhomology were demonstrated between 6 x 106 daltons and 108 x 106 daltons inthe DNAs. The structural organization of herpesvirus pan DNA was similar to theformat of Epstein-Barr virus and herpesvirus papio DNAs. The DNA consists oftwo domains of largely unique sequence complexity, a segment Us of 9 x 106daltons and a segment UL of 88 x 106 daltons. Us and UL are separated by avariable number of tandem repetitions of a sequence IR (2 x 106 daltons). Therewas homology between DNA which mapped at 26 to 28 x 106 daltons and 93 to 95x 106 daltons in UL. The terminal reiteration component, TR, of herpesvirus panDNA and sequences which mapped to the left of 6 x 106 daltons and to the right of108 x 106 daltons had no detectable homology with the corresponding regions ofEpstein-Barr virus DNA.

Herpesvirus pan (HVPan), Herpesvirus papio(HVPapio), and Epstein-Barr virus (EBV) areendemic in their respective primate species:chimpanzees, baboons, and humans (6-9, 13,17, 22, 24, 36). The viral capsid, membrane, andearly antigens specified by each of these virusesextensively cross-react with antisera from theheterologous primate species (6-9, 13, 22, 24,36). The host range of each of these viruses invitro is limited to primate B lymphocytes inwhich the virus induces growth transformationand expression of an intranuclear antigen. Al-though limited cross-reactions have been detect-ed, the intranuclear antigens specified by each ofthese viruses have major non-cross-reactive de-terminants (8, 9, 27, 32).

Previous studies of HVPan, HVPapio, andEBV DNAs indicated that (i) HVPapio andHVPan have 40% homology to EBV DNA (6-9),(ii) the sizes ofEBV and HVPapio DNAs are 100to 114 x 106 daltons (3, 4, 10, 14-16, 18-23, 29,30), (iii) EBV and HVPapio DNAs have similarorganizational formats: a variable number oftandem direct repeats of a 500 to 600-base pairsequence (TR) at both ends of the DNA, avariable number of tandem direct repeats of a3,071-base pair sequence IR which divides thegenome into a short unique region Us and a

t Deceased.

long unique region UL, and the duplication ofsequences which map at 26 to 28 and 93 to 95 x10° daltons in UL (3, 4, 10-12, 14-16, 18, 21, 23,30), and (iv) labeled fragments of EBV DNAthrough the entire genome hybridize to frag-ments at identical map positions in HVPapioDNA (16). Regions of nonhomology betweenEBV and HVPapio DNAs are confined to thetermini and to DNA which maps at 54 to 59 x106 daltons (16).The purpose of the experiments reported here

is (i) to determine whether HVPan DNA is moreor less related to HVPapio DNA than to EBVDNA, (ii) to determine whether HVPan DNAshares a common structural architecture withEBV and HVPapio DNAs, and (iii) to deriverestriction endonuclease maps for HVPan DNA.Based on the previous observation that eachcloned restriction enzyme fragment of EBVDNA is a specific probe for sequences at thecorresponding map position in HVPapio DNA(16), we used the clone library of recombinantEBV DNA fragments to identify and map frag-ments of the HVPan genome. Restriction endo-nuclease maps of HVPan DNA were construct-ed from the sizes ofHVPan DNA fragments andtheir linkage relationships. The linkage relation-ships of the restriction endonuclease fragmentsof the HVPan DNA were determined by hybrid-izations with labeled EBV DNA fragment

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932 HELLER, GERBER, AND KIEFF

probes. This approach enabled us to determinethat the EBV and HVPan genomes are homolo-gous and colinear over almost their entirelengths.

MATERIALS AND METHODSCell culture, virus purification, and preparation of

viral DNA. HVPapio isolates Ba65 and Ba74 are virus-producing baboon lymphoid cells established fromcirculating lymphocytes of a normal baboon, Papiocynocephalus (8). HVPan 1 (chimp, 8,9) and HVPan 2(Austen, gift of H. Rabin, Frederick Cancer ResearchCenter) are virus-producing chimpanzee lymphoid celllines. The procedure for purification of virus from thesupernatant media of cell cultures and the preparationof viral DNA from dextran-banded virus have beendescribed (29). HVPan 1 cell cultures produced verylimited amounts of extracellular virus. Therefore,HVPan 1 viral DNA was partially purified from asodium dodecyl sulfate-proteinase K lysate of HVPan1 cells by CsCl density equilibrium sedimentation aspreviously described (14). Recombinant EBV B95-8 orW91 DNA BamHI fragments cloned in pBR322 andEcoRI fragments cloned in Charon 4A or MUA-3 weregifts from T. Dambaugh, N. Raab-Traub, and M.Hummel (3, 30).

Labeling and hybridization ofDNAs. The proceduresfor radioactive labeling of DNA by nick translation,restriction endonuclease digestion of DNA, separationof DNA fragments by agarose gel electrophoresis,Southern blotting, hybridization of labeled DNAs tonitrocellulose filters, and the analysis of DNA-DNAreassociation kinetics with S1 nuclease have beenpreviously described (10, 11, 15, 28, 29). Probe DNAswere hybridized to Southern blots in 1.0 M Na+ at74°C. DNA-DNA reassociation in solution was per-formed in 1.0 M Na+-5% formamide at 74°C.

RESULTSHomology between HVPan and HVPapio

DNAs. Hybridization of nick translated EBVDNA with excess unlabeled HVPan or HVPapioDNAs renders about 40% of the labeled EBVDNA resistant to degradation by the single-strand specific exoendonuclease Si. This indi-cates that 40% of the nucleotides of HVPan andHVPapio are in highly conserved oligo-polynu-cleotide sequences (39). To determine whetherHVPan and HVPapio DNAs are more closelyrelated to each other than to EBV DNA, HVPa-pio DNA was extracted from dextran-bandedvirus, purified by CsCl density equilibrium sedi-mentation, labeled by nick translation, dena-tured, and incubated with an excess of dena-tured HVPan, EBV, or HVPapio DNAs understringent hybridization conditions (25°C belowthe Tm of EBV DNA). The concentration ofeach unlabeled DNA was adjusted so that theinitial rates of hybridization of the unlabeledDNAs to labeled HVPapio DNA were similar.HVPan and EBV DNAs hybridized to 42 and36% of the HVPapio probe, respectively (Fig.1), at 5 x COt50 (18 h) of reassociation. There-

'or

0

C/co .5

o

- (0) HVPon I(a) P3HR -I(0) HVPopio (Bo65)

- (C) HVPopb (Bo74)

10 10 lo' 10'cot

FIG. 1. Reassociation of 32P-labeled HVPapio(Ba65) DNA (2.5 ng/ml) in the presence of 20 .ig ofEBV (P3HR-1) DNA purified from virus per ml,HVPan 1 DNA purified from intracellular DNA, orHVPapio (Ba74 or Ba65) DNAs purified from virus.The fraction of HVPapio (Ba65) probe DNA remainingsingle stranded (C/CO) was determined at the indicatedCot (moles of nucleotide per liter -l * s) with S1nuclease. C/CO in the reassociation has been correctedfor the self-association of the probe in the presence ofsalmon sperm DNA. Cot values have been corrected toequivalent Cot in 0.18 M Na'.

fore, EBV, HVPapio, and HVPan DNAs are allsimilarly related to each other.

Restriction endonuclease fragments of HVPanDNAs. HVPan 1 DNA was separated from in-fected cellular DNA by isopycnic banding inneutral CsCl. HVPan 2 DNA was extracted fromHVPan 2 virions which were purified from theextracellular fluid of HVPan 2-infected cells.The sizes of the EcoRI fragments of HVPan 1and HVPan 2 DNAs were similar, except thatEcoRI fragment G of HVPan 1 DNA was 7.0 x106 daltons, whereas EcoRI fragment G ofHVPan 2 DNA was 8.2 x 106 daltons (Table 1).The sum of the sizes of the fragments of HVPan1 DNA was 105 to 115 x 106 daltons. The largestfragments of HVPan 1 DNA were underrepre-sented, probably as a consequence of shearingduring extraction with cell DNA and repeatedcycles of isopycnic centrifugation. Most of theremaining HVPan 1 and HVPan 2 DNA frag-ments were represented in equimolar propor-tions in ethidium bromide-stained gels and inautoradiograms of Southern blots hybridizedwith labeled HVPan 1 DNA (Fig. 2 and 3).

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DNA OF HERPESVIRUS PAN 933

TABLE 1. Size of HVPan DNA restriction endonuclease fragmentsHVPan 1 HVPan 2

XbaI BamHI EcoRI EcoRI

Fragment Mol wt x 106 Fragment Mol wt x 106 Fragnent Mol wt x 106 Fragment Moi wt x 106

A 19 A 16 A 18 A 24B 17 B 6.1 B 14 B 13.5C 14 C 5.9 C 13.5 C 13Dhet 13.5 D 4.8 D 11 D 10E 13.5 E 4.8 E 9.4 E 9.2F 8.2 F 4.5 F 8.0 F 8.2G 6.8 G 4.5 G 7.0 G 8.2H 5.4 H 4.0 H 5.5 H 5.5I 5.1 I 3.5 I 5.1 I 5.2J 1.7 J 3.2 J 4.0 J 4.1

K1, K2, K3 3.2 K 2.5 K 2.5L 3.0 L 2.1 L 2.1M 2.8 M 2.0 M 2.0N 2.5 N 1.1 N 1.10 2.2P 2.2Q 1.8R 1.8S 1.6T 1.5U 1.4V 1.2W 1.1X 0.95Y 0.9Z 0.8a 0.7b 0.7c 0.7d 0.6

e, f 0.5

Submolar fragments can be clearly discerned inthe autoradiograms of EcoRI fragments ofHVPan 1 (Fig. 2A) below the EcoRI-A fragmentand as pairs of fragments between EcoRI-J andEcoRI-K. The submolar fragments belowEcoRI-A and HindIII-B (Fig. 2) differ from eachother by increments of 2 x 106 daltons, suggest-ing that HVPan 1 DNA may contain a variablenumber of repeats of an IR of 2 x 106 daltons ashas been previously observed in EBV andHVPapio DNAs (11, 14, 15). Labeled EBV IRprobe specifically identified the IR componentsof HVPan DNA in Southern blot hybridizations(Fig. 2).The presence of tandem repeats of IR in

HVPan DNA was confirmed by testing severalrestriction endonucleases to determine whetherany cut once within the putative IR sequence,thereby generating a fragment of 2 x 106 daltonspresent in excess molar ratio relative to otherHVPan DNA fragments. XhoI, BgII, SstI, andSstII cleavages of HVPan DNA yielded frag-ments of 2 x 106 daltons in excess molar ratio toother HVPan 1 DNA fragments. The supermolar

fragments of 2 x 106 daltons were identical insize to the IR of EBV DNA and hybridizedextensively to it (Fig. 2C). Labeled EBV IRidentified two additional HVPan SstII frag-ments. These fraginents probably contain thejunctions of HVPan IR with the Us and ULregions and would therefore consist of the rightend of Us and IR sequences to the left of theSstII site and the left end of UL and IR se-quences to the right of the SstII site.

Restriction endonudease maps ofHVPan DNA.Replicate Southern blots were made from agar-ose gels of EcoRI, XbaI, and BamHI fragmentsof HVPan 1 DNA. Labeled cloned EBV DNAfragments were hybridized to the blots to identi-fy the EcoRI, XbaI, and BamHI fragmentswhich contain sequences related to the EBVprobe fragments (Fig. 3). The sizes of theHVPan DNA fragments were estimated from themigration of the fragments relative to bacterio-phage lambda DNA fragment standards (36,Table 1). These data establish a linkage map ofthe EcoRI, XbaI, and BamHI restriction endo-nuclease fragments of HVPan 1 DNA (Fig. 4).

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934 HELLER, GERBER, AND KIEFF

A Eiol.-PHtP

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FIG. 2. Restriction endonuclease profiles and detection of an internal reiteration component in HVPan DNA.The cloned internal reiteration component of EBV DNA, BamHI-V (V), was labeled by nick translation andhybridized to nitrocellulose paper containing the separated EcoRI (A) and HindIII (B) restriction enzymefragments of HVPan 2 DNA. The blot was rehybridized with labeled HVPan 2 DNA (TP) to show all of thefragments. The sizes in megadaltons of intact and EcoRI fragments of bacteriophage lambda DNA are indicated.(C) Labeled EBV BamHI-V-cloned plasmid DNA was hybridized to a Southern blot of an agarose gel containingHVPan 1 CsCI-enriched intracellular DNA digested with the indicated restriction enzymes and recombinantEBV BamHI V-pBR322 DNA cleaved with BamHI. The HVPan 1 DNA fragments identical in size to EBVBamHI-V and identified by V probe are assumed to represent the HVPan 1 DNA internal reiteration (IR).

V -

For example, labeled EBV BamHI-U hybridizedto HVPan 1 EcoRI-H, XbaI-B, and BamHI-T,-D, and -E. These EcoRI, XbaI, and BamHIfragments must therefore overlap in the HVPangenome. HVPan BamHI-T shares sequenceswith labeled EBV BamHI-P probe. This probefragment hybridized to HVPan EcoRI-J only.Therefore, EcoRI-J maps next to EcoRI-H. Theremaining fragment linkage relationships wereestablished in a similar way. The HVPan 1 DNArestriction enzyme fragment map is presented inFig. 4.The HVPan DNA fragments are shown with

the homologous EBV probe fragment (Fig. 3 and4). Most EBV probes hybridized to only a singleregion of the HVPan genome. EBV DNA frag-ments containing sequences duplicated at twolocations in UL of EBV, BamHI-H and BamHI-Bi (Fig. 4; [30]) are an exception and hybridizeto both regions in HVPan DNA. The duplicationof sequences in UL is therefore a conservedfeature between EBV and HVPan DNAs. EBVBamHI-H and BamHI-B1 probes also hybrid-ized to the same submolar HVPan 1 EcoRI andBamHI fragments, indicated by asterisks in Fig.3. The fragments do not appear to contain se-quences homologous to other regions of EBV

DNA. Similar submolar fragments have beendetected in Southern blots of intracellular EBVDNAs (14).The labeled EBV terminal fragment EcoRI-D,

-I-J het, which contains DNA from the leftmostregions of Us and the rightmost portions of ULand TR, hybridized to HVPan DNA fragmentswhich mapped to the left of 108 x 106 daltons.This probe identified only one terminal hetero-geneous fragment in HVPan 1 DNA, the XbaI-Dhet fragment. This hybridization to XbaI-D hetmost likely reflects sequence relatedness whichmaps at 105 to 108 x 106 daltons in the EBV andHVPan genomes (Fig. 4). There was no detect-able homology between the TR components ofEBV and HVPan DNAs. To further examine thetermini ofHVPan DNA, the HVPapio HindIII-Rhet fragment of 1 x 106 daltons, which containsthe TR of HVPapio DNA (15, 16), was labeledby nick translation and hybridized to blots ofHVPan and EBV DNA fragments. This probedid not hybridize to the TR of HVPan or EBVDNAs (data not shown). Other EBV Us probesdid not hybridize to HVPan DNA fragmentswhich mapped to the left of 6 x 106 daltons. TheTR and immediately adjacent unique sequencesin Us and UL were the only regions of nonho-

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DNA OF HERPESVIRUS PAN 935

mology between EBV and HVPan 1 DNAs.Aside from this exception and the duplication ofsequences in UL, each labeled EBV probe hy-bridized to the equivalent region of the HVPangenome.

DISCUSSIONThe B-lymphotropic herpesviruses related to

EBV have a similar organizational format whichdistinguishes the group from the other herpes-viruses (3, 4, 10-12, 14-16, 18, 23, 30). Thecommon DNA structure consists of two compo-nents of unique DNA, a Us of 9 x 106 daltonsand a UL of 88 x 106 daltons which are separat-ed by a variable number ofdirect tandem repeatsof an IR of 2 x 106 daltons. The linear genome isbounded by a variable number of repeats of a TRof 3 to 3.5 x 105 daltons at both ends. The DNAsof HVPan, HVPapio, and EBV are related andcolinear, except for the termini and the region 54to 59 x 106 daltons which is nonhomologous inEBV and HVPapio DNAs (15, 16). Homologybetween sequences mapping at 26 to 28 and 93 to95 x 106 daltons is conserved in the three DNAs(15, 16, 30). From the extent of conservation offormat among these three DNAs, it is likely thatthe B-lymphotropic herpesviruses of gorillas andorangutans will share this DNA structure, par-ticularly since these viruses are all closely relat-ed by immunological criteria (7, 9, 26, 31).There is nucleotide sequence conservation

and some divergence among HVPan, HVPapio,and EBV DNAs, since only 40%o of the se-quences are able to form oligo-polynucleotidebase pairs under stringent reassociation condi-tions (6, 9). This is a minimum estimate of DNAsequence conservation among these DNAs. AtTm - 25°C, regions of only partial homologyamong the DNAs could form heteroduplexeswhich would be S1 nuclease sensitive owing tointerspersed unmatched base pairs (39). Theconserved sequences are colinear among thegenomes and are distributed across almost thefull length of the DNAs (16). The colinear relat-edness of EBV, HVPan, and HVPapio DNAsover almost the entire length of the moleculesindicates that these primate viruses evolvedfrom a common ancestor.EBV and HVPan DNAs share the same de-

gree of homology to HVPapio DNA, and HVPa-pio and HVPan DNAs share the same degree ofhomology to EBV DNA. This suggests that thethree viruses evolved from a common parent atabout the same time. Two evolutionary modelsare suggested: (i) a vertical model in which theevolution of the viruses was contemporary withthe evolution of the host primate species or (ii) ahorizontal model in which a common progenitorvirus spread among the host species at some

point after the onset of species divergence.Nucleotide sequence analysis of viruses fromprimates which share common and diverse evo-lutionary branch points should discriminate be-tween these two hypotheses. Studies of nucleicacid homologies among the nonrepeated se-quences of primate DNAs have provided a mea-sure of the evolutionary relationships amongapes and Old World monkeys (1). Data on theextent of reassociation of nonrepeated cellularDNAs and the thermal stability of these DNAhybrids suggest that humans are closely relatedto the chimpanzee and gorilla and progressivelymore distantly related to other apes and OldWorld monkeys, including baboons (1). Thesimilar genetic relatedness among EBV,HVPan, and HVPapio DNAs is not consistentwith the evolutionary relationships of the hostspecies. These data therefore favor the horizon-tal spread of an ancestral virus among the pri-mate host species after the onset of speciesdivergence.The extent to which structural organization

and nucleic acid sequence are conserved amongHVPapio, HVPan, and EBV DNAs suggests theoperation of selective pressures which maintainthe genetic relatedness of these genomes. Someof the constraints on the position and sequenceof a region of DNA within a genome arise frommultiple utilization of the same DNA sequencefor overlapping or complementary codons, acommon leader sequence for several mRNAs, ora common control region for several genes.These phenomenon are well documented insmaller viral genomes (e.g., adenovirus; 5, 34,35). Another constraint which may have acted tolimit the divergence of the genomes is mainte-nance of sufficient sequence homology and com-patible structural genetic organization to permitviable reassortments through homologous re-combination. The latter may be selective in theevolution of bacteriophage whose hosts sharea common ecological habitat (2). It may be lessrelevant to the primate herpesviruses, sincetheir hosts do not naturally interact. Further-more, the high prevalence of homologous infec-tion and immunity makes chance infection withthe heterologous agent unlikely.The consistent lack of homology among the

terminal reiteration sequences of EBV, HVPan,and HVPapio DNAs contrasts with the exten-sive homology among the IRs and the duplica-tion of related sequences in UL of these ge-nomes. IR encodes abundant polyadenylatedcytoplasmic RNAs in latently infected growth-transformed cells (19, 20, 38). No mRNAs havebeen mapped to TR. The evolution of TR mightbe less constrained if it is a structural elementwhich facilitates the circularization of the linearDNA after infection (25).

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936 HELLER, GERBER, AND KIEFF

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VOL. 41, 1982 DNA OF HERPESVIRUS PAN 937

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FIG. 4. Restriction endonuclease fragment linkage map of HVPan 1 DNA and summary of cross hybridiza-tions between EBV and HVPan 1 DNAs. HVPan 1 DNA EcoRI (A), XbaI (B), and BamHI (C) fragments havebeen aligned with the EBV-cloned fragments (D) which were hybridized to HVPan DNA fragments in this study.The sizes of the HVPan 1 DNA fragments were estimated from their migration relative to bacteriophage lambdaDNA EcoRI and HindIII fragments in agarose gels. The homology data also represent a map ofEcoRI, XbaI, andBamHI restriction enzyme sites in HVPan 1 DNA. Since the terminal reiteration (TR) components and uniquesequences to the left of 6 x 106 daltons and to the right of 108 x 106 daltons in EBV and HVPan 1 DNAs do nothave detectable sequence homology, the uncertainty in the location of restriction enzyme sites in these regions ofHVPan 1 DNA is represented by a dashed line. The general structure of the EBV and HVPapio genomes isshown in the bottom panel.

ACKNOWLEDGMENTSThe HVPan 2 cell line was contributed by H. Rabin of the

Frederick Cancer Research Center. We thank H. Swift forhelpful discussion.

This research was supported by Public Health Servicegrants CA 19264 and CA 17281 from the National CancerInstitute and by American Cancer Society grant ACS MV 32F.M. H. is a predoctoral student supported by Public HealthService training grant AI-07099 from the National Institutes ofAllergy and Infectious Disease. E. K. is a Faculty ResearchAwardee of the American Cancer Society.

LITERATURE CITED1. Benveniste, R. E., and G. J. Todaro. 1976. Evolution of

type C viral genes: evidence for an Asian origin of man.Nature (London) 261:101-108.

2. Botstein, D. 1980. A theory of modular evolution forbacteriophage. Ann. N.Y. Acad. Sci. 354:484-491.

3. Dambaugh, T., C. Belsel, M. Hummel, W. King, S.Fennewald, A. Cheung, M. Heller, N. Raab-Traub, and E.Kieff. 1980. Epstein-Barr virus DNA. VII. Molecularcloning and detailed mapping ofEBV (B95-8) DNA. Proc.Natl. Acad. Sci. U.S.A. 77:2999-3003.

4. Dambaugh, T., N. Raab-Traub, M. Heller, C. Beisel, M.Hummel, A. Cheung, S. Fennewald, W. King, and E. Kieff.1980. Variations among isolates of Epstein-Barr virus.Ann. N.Y. Acad. Sci. 602:711-719.

5. Darnell, J., E. Ziff, J. Nevins, and M. Wilson. 1980.Variety in the mechanisms of adenovirus gene control.Ann. N.Y. Acad. Sci. 354:466-471.

6. Falk, L., F. Deinhardt, M. Nonoyama, L. Wolfe, and C.Bergholz. 1976. Properties of a baboon lymphotropicherpesvirus related to Epstein-Barr virus. Int. J. Cancer18:798-807.

7. Gerber, P., and S. M. Birch. 1967. Complement-fixingantibodies in sera of human and non-human primates toviral antigens derived from Burkitt's lymphoma cells.Proc. Natl. Acad. Sci. U.S.A. 58:478-484.

8. Gerber, P., S. S. Kalter, G. Schidlovsky, W. D. Peterson,Jr., and M. D. Daniel. 1977. Biologic and antigeniccharacteristics of Epstein-Barr virus-related herpes-viruses of chimpanzees and baboons. Int. J. Cancer20:448-459.

9. Gerber, P., R. Pritchett, and E. Kieff. 1976. Antigens andDNA of a chimpanzee agent related to Epstein-Barr virus.J. Virol. 19:1090-1100.

10. Given, D., and E. Kieff. 1978. DNA of Epstein-Barr virus.IV. Linkage map for restriction enzyme fragments of theB95-8 and W91 strains of EBV. J. Virol. 28:524-542.

11. Given, D., and E. Kieff. 1979. DNA of Epstein-Barr virus.VI. Mapping of the internal tandem reiteration. J. Virol.31:315-324.

12. Given, D., D. Yee, K. Griem and E. Kieff. 1979. DNA ofEpstein-Barr virus. V. Direct repeats at the ends of EBVDNA. J. Virol. 30:852-862.

13. Goldman, N., H. C. Landon, and J. I. Resbher. 1968.Fluorescent antibody and gel diffusion reactions of humanand chimpanzee sera with cells cultured from Burkitttumors and normal chimpanzee blood. Cancer Res.28:2489-2495.

14. HeUler, M., T. Dambaugh, and E. Kieff. 1981. Epstein-Barr virus DNA. IX. Variation among viral DNAs. J.Virol. 38:632-648.

15. HeUer, M., P. Gerber, and E. Kieff. 1981. Herpesviruspapio DNA is similar in organization to Epstein-Barr virusDNA. J. Virol. 37:698-709.

16. HeUer, M., and E. Kieff. 1981. Colinearity between theDNAs of Epstein-Barr virus and herpesvirus papio. J.Virol. 37:821-826.

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17. Henle, W., and G. Henle. 1980. Epidemiologic aspects ofEpstein-Barr virus (EBV)-associated diseases. Ann. N.Y.Acad. Sci. 354:326-331.

18. Kieff, E., D. Given, A. L. T. Poweli, W. King, T. Dam-baugh, and N. Raab-Traub. 1979. Epstein-Barr virus:structure of the viral DNA and analysis of viral RNA ininfected cells. Biochim. Biophys. Acta Rev. 560:355-373.

19. King, W., A. L. Thomias-Powell, N. Raab-Traub, M.Hawke, and E. Kieff. 1980. Epstein-Barr virus RNA. VI.Viral RNA in a restringently infected, growth-trans-formed cell line. J. Virol. 36:506-518.

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