臺灣博碩士論文加值系統

中文摘要：
　　胸腺核激(Thymidine kinase, TK)是DNA合成的救助途徑中，將胸腺核磷酸化為胸腺核單磷酸的酵素。本實驗室前已將EB病毒TK的cDNA成功轉殖至表現質體pET3cp*，並在大腸菌中成功的表現大量具有活性的EB病毒TK重組蛋白，之後更將TK基因再轉殖至表現質體pET21c，在大腸菌中表現出在羧基端帶有連續組織胺酸殘基序列的TK重組蛋白，並以鎳離子管柱進行了純化。
　　對於EB病毒TK蛋白質結構與反應機制的認識，長久以來來自於對單純疹病毒(herpes simplex virus, HSV)TK蛋白質的研究，雖然兩者的胺基酸序列極為相似，但在生化性質上仍不盡相同。HSV TK蛋白質的Asp162被推測與鎂離子的定位有關。1996年Black等人將其突變為glutamine，結果突變株TK蛋白質的活性大幅下降，對鎂離子的親和力卻沒有明顯改變。為了證明EB病毒TK蛋白質上的相對位置，Asp392，是否具有相似的功能，本實驗將其亦突變為glutamine，並利用鎳離子管柱純化突變株TK蛋白質後分析其特性。結果與HSV TK蛋白質的研究相似，活性明顯下降，但與鎂離子的結合能力卻與HSV TK蛋白質的研究結果不同，突變株EB病毒TK蛋白質的活性似乎受到鎂離子的抑制。
　　藉由冷凍-解凍再過濾的過程，發現了藉由pET21c表現載體所表現的TK重組蛋白質有強烈的凝聚現象，利用pET3cp*表現的TK重組蛋白質則無此現象，推測可能是TK重組蛋白末端的連續組織胺酸殘基序列造成了較容易凝聚現象。
鼻咽癌是台灣十大癌症死因之一。而抗EB病毒DNA分解抗體是一項檢測鼻咽癌的有效指標。由於鼻咽癌的發生與遺傳及環境因素有關，本實驗篩檢了51個含有鼻咽癌病患的家族（包含了49名病患與257名家屬），141名電信局員工與81名報社員工。近來發現胃癌的發生也與感染EB病毒有關，因此本實驗也針對213名胃癌病患與84名非胃癌病患進行篩檢。
實驗結果顯示鼻咽癌病患家屬的抗EB病毒DNA分解抗體陽性率是9.7%，將近正常人 (約5%) 的兩倍。而針對鼻咽癌病患所測得的陽性率 (65.3%) 雖然低於以往實驗室所測得的值 (80~90%)，但是其中大多數呈現陽性的患者具有高效價的抗EB病毒DNA分解抗體。檢測電信局員工的結果，其陽性率(5.6%) 與正常人並無明顯差異。但在檢測報社員工的結果中卻發現了明顯偏高的陽性率 (25.9%)。針對胃癌病患所測得的陽性率 (16.9%) 與本實驗室已往檢測其他種癌症病患的陽性率 (10~30%) 近似，但在檢測非胃癌患者時卻發現了不尋常的高陽性率，推測可能是由於實驗誤差所導致的假陽性率。

Abstract:
Thymidine kinase (TK) is the enzyme that phosphorylates thymidine (TdR) to thymidine monophosphate (TMP) during the salvage pathway of DNA synthesis. In this laboratory, a recombinant Epstein-Barr virus (EBV) TK cDNA, pET3cp*-TKB1B, has been previously constructed. The recombinant TK protein, 3cp*-TK, has also been successfully expressed in E. coli system with high quantity and high enzyme activity. The EBV TK coding sequence was also subcloned into pET21c and another recombinant EBV TK protein, 21c-TK, with 6 histidine residues at the carboxyl terminus was obtained, allowing the purification of TK protein through a nickel affinity column.
The knowledge of EBV TK protein structure and its reaction mechanism mostly came from researchs on herpes simplex virus (HSV) TK. Though sharing lots of sequence similarities, these two enzymes are still somehow different in biochemical properties. Asp162 of HSV TK has been suggested to function for coordinating Mg2+. To find out if the analog of Asp162 in EBV TK, Asp392, plays the same role, an EBV TK mutant with a glutamine substituted Asp392 was generated and named 21c-TK(D392Q). Both 21c-TK(D392Q) and 21c-TK were purified via nickel affinity column. As a result, 21c-TK(D392Q) exhibited similar properties as the HSV TK D162Q mutant protein except that its activity seemed to be inhibited by Mg2+ .
Through the freezing-thawing process, purified 21c-TK and 21c-TK(D392Q) aggregated seriously. To discover the cause of aggregation, the tendency to aggregation for TK proteins was compared between 21c-TK and 3cp*-TK. After
35,000 rpm centrifugation for 1 hour and filtration, 21c-TK still aggregated much more obviously than 3cp*-TK did. The results suggested that the extra His-tag at the carboxyl terminus of 21c-TK might contribute the easier aggregation.
Nasopharyngeal carcinoma (NPC) is one of the ten most common cancers in Taiwan. Antibody to EBV specific DNase has been proved to be a useful marker for early detection of NPC. NPC development is affected by genetic factors and environmental factors. Therefore, family members from 51 NPC patients (including 49 patients and 257 relatives), 141 employees of a telegraph office and 81 employees of a newspaper company were examined. Gastric cancer is lately proved associated with EBV infection. Thus, sera from 213 gastric carcinoma patients and 84 patients without gastric carcinoma were also included.
The result shows the anti-EBV DNase positive rate of NPC patients’ families is nearly twice the value of healthy group examined previously (9.7% vs 5%). The positive rate of NPC patients is lower than previous results (65.3% vs 80~90%), yet most of the positive individuals have high level anti-EBV DNase. The positive rate of employees of a telegraph office shows no difference with healthy group positive rate (5.6% vs 5%), whereas a significant high positive rate (25.9%) was detected in employees of a newspaper company. Gastric carcinoma patients examined have similar positive rate with other cancer patients examined previously (16.4% vs 10~30%). The non-gastric carcinoma patients have an unusual high positive rate (50%), possibly due to examination error.

目 錄
中文摘要………………………………………. I
英文摘要……………………………………… II
縮寫表………………………………………. III
序論……………………………………………. 1
實驗材料與方法…………………………….. 18
實驗結果…………………………………….. 35
討論………………………………………….. 43
圖表………………………………………….. 51
實驗步驟…………………………………….. 76
參考文獻…………………………………….. 96

Archard L, Peters L, Behan P, Doyle D, Bowles N (1988): Detection of Epstein-Barr virus DNA in muscle from patients with the postviral chronic fatigue syndrome, myalgic encephalomyelitis. Journal of Experimental and Clinical Cancer Research 7: 142-145.
Baer R, Bankier AT, Biggin MD, Deininger PL, Farrell PJ, Gibson TJ, Hatfull G, Hudson GS, Satchwell SC, Seguin C, Tuffnell PS, Barrell BG (1984): DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature (London) 310: 207- 211.
Balasubramaniam NK, Veerisetty V, Gentry GA (1990): Herpesviral deoxythymidine kinases contain a site analogous to the phosphoryl-binding arginine-rich region of porcine adenylate kinase; comparison of secondary structure predictions and conservation. Journal of General Virology 71: 2979-2987.
Black ME, Loeb ZA (1993): Identification of important residues within the putative nucleoside binding site of HSV-1 thymidine kinase by random sequence selection :analysis of selected mutants in vitro. Biochemistry. 32: 11618-11626.
Black ME, Rechtin TM, Drake RR (1996): Effect on substrate binding of an alteration at the conserved aspartic acid-162 in herpes simplex virus type 1 thymidine kinase. Journal of General Virology 77:1521-1527.
Brown DG, Visse R, Sandhu G, Davies A, Rizkallah PJ, Melitz C, Summers WC, Sanderson MR (1995): Crystal structures of the thymidine kinase from herpes simplex virus type 1 in complex with deoxythymidine and gancyclovir. Nature Structural Biology 2: 876-880.
Burke AP, Yen TS, Shekitka KM, Sobin LH (1990): Lymphoepithelial carcinoma of the stomach with Epstein-Barr virus demonstrated by polymerase chain reaction. Modern Pathology 3: 377-380.
Chang YR (1994): I. Purification of Epstein-Barr virus thymidine kinase and preparation of monoclonal antibodies. II. Nucleotide sequencing and generation of carboxyl terminus mutants. Thesis for Master Degree, Graduate Institute of Microbiology, College of Medicine, National Taiwan University.
Chen CL, Sadler RH, Walling DM, Su JJ, Hsieh HC, Raab-Traub N (1993a): Epstein-Barr virus (EBV) gene expression in EBV-positive peripheral T-cell lymphomas. Journal of Virology 67: 6303-6308.
Chen JY, Chen CJ, Liu MY, Cho SM, Hsu MM, Lynn TC, Shieh T, Tu SM, Beasley RP, Hwang LY, Lee HH, Kou SL, Yang CS (1989): Antibody to Epstein-Barr virus-specific DNase as a marker for field survey of patients with nasopharyngeal carcinoma in Taiwan. Journal of Medical Virology 27:269-273.
Chen JY, Chen CJ, Liu MY, Cho SM, Hsu MM, Lynn TC, Shieh T, Tu SM, Lee HH, Kuo SL, Lai MY, Hsieh CY, Hu CP, Yang CS (1987): Antibodies to Epstein-Barr virus-specific DNase in patients with nasopharyngeal carcinoma and control groups. Journal of Medical Virology 23:11-21.
Chen JY, Hwang LY, Beasley RP, Chien CS, Yang CS (1985a): Antibody response to Epstein-Barr-virus-specific DNase in 13 patients with nasopharyngeal carcinoma in Taiwan: a retrospective study. Journal of Medical Virology 16:99-105.
Chen JY, Liu MY, Chen CJ, Hsu MM, Tu SM, Lee HH, Kuo SL, Yang CS (1985b): Antibody to Epstein-Barr virus-specific DNase as a marker for the early detection of nasopharyngeal carcinoma. Journal of Medical Virology 17:47-49.
Chen JY, Liu MY, Hsu TY, Cho SM, Yang CS (1993b): Use of bacterially-expressed antigen for detection of antibodies to the EBV-specific deoxyribonuclease in sera from patients with nasopharyngeal carcinoma. Journal of Virological Methods 45:49-66.
Chen JY, Liu MY, Lynn TC, Yang CS (1982): Antibody to Epstein-Barr virus-specific DNase in patients with nasopharyngeal carcinoma. Chinese Journal of Microbiology and Immunology 15:255-261.
Chen ST, Estes JE, Huang ES, Pagano JS (1978): Epstein-Barr virus-associated thymidine kinase. Journal of Virology 26: 203-208.
Cheng YC, Chen JY, Glaser R, Henle W (1980): Frequency and levels of antibodies to Epstein-Barr virus-specific DNase are elevated in patients with nasopharyngeal carcinoma. Proceedings of the National Academy of Sciences USA 77:6162-6165.
Chevallier-Greco A, Manet E, Chavrier P, Mosnier C, Daillie J, Sergeant A (1986): Both Epstein-Barr virus (EBV) encoded transacting factors, EB1 and EB2, are required to activate transcription from an EBV early promoter. EMBO Journal 5: 3243-3249.
Clough W (1979): Deoxyribonuclease activity found in Epstein-Barr virus producing lymphoblastoid cells. Biochemistry 18: 4517-4521.
Countryman J, Miller G (1985): Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA. Proceedings of the National Academy of Sciences USA 82: 4085-4089.
Countryman J, Jenson H, Rudi S, Wolf H, Miller G (1987): Polymorphic proteins encoded within BZLF1 of defective and standard Epstein-Barr viruses disrupt latency. Journal of Virology 61: 3672-3679.
Cox MA, Leahy J, Hardwick JM (1990): An enhancer within the divergent promoter of Epstein-Barr virus responds synergistically to the R and Z transactivators. Journal of Virology 64: 313-321.
Darby G, Larder BA, Inglis MM (1986): Evidence that the ‘active center ’of the herpes simplex virus thymidine kinase involves an interaction between three distinct regions of the peptide. Journal of General Virology 67: 753-758.
De Turenne-Tessier M, Ooka T, Calender A, De The G, Daillie J (1989): Relationship between nasopharyngeal carcinoma and high antibody titers to Epstein-Barr virus specific thymidine kinase. International Journal of Cancer 43: 45-48.
Dolyniuk M, Pritchett R, Kieff E (1976): Proteins of Epstein-Barr virus. I. Analysis of the polypeptides of purified enveloped Epstein-Barr virus. Journal of Virology 17: 935-949.
Epstein MA, Achong BG, Barr YM (1964): Virus particles in cultured lymphoblasts from Burkitt’s lymphoma. Lancet 1: 702-703.
Fahraeus R, Hu LF, Ernberg I, Finke J, Rowe M, Klein G, Falk K, Nisson E, Yadav M, Busson P,Tursz T,Kallin B (1988):Expression of Epstein-Barr virus-encoded proteins in nasopharyngeal carcinoma. International Journal of Cancer 42: 329-398.
Fetzer J, Folkers G, Muller I, Keil GM (1993): Site-directed mutagenesis in the active site of the herpes simplex virus type 1 thymidine kinase gene. Virus Genes 7: 205-209.
Folkers G, Trumpp-Kallmeyer S, Gutbrod O, Krickl S, Fetzer J, Keil GM (1991): Computer-aided active-site-directed modeling of the herpes simplex virus 1 and human thymidine kinase. Journal of Computer Aided Molecular Design 5: 385-404.
Gabhann PM, Sugawara K, Ito Y (1984): Characterization of Epstein-Barr virus—related thymidine kinase induced in nonproducer cells by superinfection or chemical treatment. Intervirology 21: 104-109.
Gentry GA (1992): Viral thymidine kinase and their relatives. Pharmacology and Therapeutics 54: 319-355.
Glaser R, Ogino T, Zimmerman J Jr., Rapp F(1973): Thymidine kinase activity in Burkitt lymphoblastoid somatic cell hybrids after induction of the EB virus. Proceedings of the Society for Experimental Biology and Medicine 142: 1059-1062.
Graessmann A, Wolf H, Bornkamn GW (1980): Expression of Epstein-Barr virus genes in different cell types after microinjection of viral DNA. Proceedings of the National Academy of Sciences USA 77: 433-436.
Hampar B, Derge JG, Martos LM, Walker JL (1971): Persistence of a repressed Epstein-Barr virus genome in Burkitt's lymphoma cells made resistant to 5-bromodeoxyuridine. Proceedings of the National Academy of Sciences USA 68: 3185-3189.
Hampar B, Derge JG, Martos LM, Walker JL (1972): Synthesis of Epstein-Barr virus after activation of the viral genome in a “virus-negative” human lymphoblastoid cell (Raji) made resistant to 5-bromodeoxyuridine. Proceedings of the National Academy of Sciences USA 69: 78-82.
Henle G, Henle W (1966): Immunofluorescence in cells derived from Burkitt’s lymphoma. Journal of Bacteriology 91: 1248-1256.
Henle G, Henle W (1976): Epstein-Barr virus-specific IgA serum antibodies as an outstanding feature of nasopharyngeal carcinoma. International Journal of Cancer 17: 1-7.
Henle G, Henle W, Diehl V (1968): Relation of Burkitt tumor-associated herpes-type virus to infectious mononucleosis. Proceedings of the National Academy of Sciences USA 59: 94-101.
Henle W, Henle G, Ho HC, Burtin P, Cachin Y, Clifford P, De Schryver A, De The G, Diehl V, Klein G (1970): Antibodies to Epstein-Barr virus in nasopharyngeal carcinoma, other head and neck neoplasms and control group. Journal of National Cancer Institute 44: 225-231.
Honess RW, O’Hare P, Young D (1982): Comparison of thymidine kinase activities induced in cells productively infected with herpesvirus saimiri and herpes simplex virus. Journal of General Virology 58: 237-249.
Hsu MM, Lynn TC, Hsieh T, Huang SC, Tu SM (1981): Factors affecting the survival of patients with nasopharyngeal carcinoma. Journal of Formosan Medical Association 80:1296-1306.
Hsu TY, Liu MW, Chang YR, Pai CY, Liu MY, Yang CS, Chen JY (1996): Function analysis of C-terminal deletion mutants of Epstein-Barr virus thymidine kinase. Journal of General Virology 77: 1893-1899.
Hsu TY, Pai CY, Shieh SM, Cho SM, Liu MY, Chen JY, Yang CS (1992): Use of antigen expressed in bacteria for detection of EBV-specific thymidine kinase antibodies in sera from patients with nasopharyngeal carcinoma. Journal of Medical Virology 38: 214-219.
Hummel M, Thorley-Lawson D, Kieff E (1984): An Epstein-Barr virus DNA fragment encodes messages for two major envelope glycoproteins (gp350/330 and gp250/200). Journal of Virology 49: 413-417.
Inglis MM, Darby G (1987): Analysis of the role of the cysteine 171 residue in the activity of herpes simplex virus type 1 thymidine kinase by oligonucleotide-directed mutagenesis. Journal of General Virology 68: 39-46.
Kallin B, Sternas L, Saemundsen AK, Luka J, Jornvall H, Eriksson B, Tao PZ, Nilsson MT, Klein G (1985): Purification of the Epstein-Barr virus DNA polymerase from P3HR1 cells. Journal of Virology 54: 561-568.
Kan SH (1997): I.Analysis of Epstein-Barr virus DNA nucleotide sequence in patients with nasopharyngeal carcinoma. II.Studies of amino terminus of Epstein-Barr virus thymidine kinase. Thesis for Master Degree, Graduate Institute of Microbiology, College of Medicine, National Taiwan University.
Kieff E (1996): Epstein-Barr virus and its replication. In Field BN and Knipes DM (eds.). Virology ( 3rd ed.). Raven Press, Ltd., Philadelphia , pp. 2343-2396.
Kit S, Dubbs DR (1969): Enzyme induction by viruses. Monographs in Virology, Vol.2.
Kit S, Kit M, Pirtle EM (1985): Attenuated properties of thymidine kinase-negative deletion mutant of pseudorabies virus. American Journal of Veterinary Research 46: 1359-1367.
Kits S, Leung WC, Lorgensen GN, Trkula D, Dubbs DR (1975): Viral-induced thymidine kinase isozymes. Progress in Medical Virology 21: 13-34.
Kit S, Sheppard M, Ichimura H, Nusinoff-Lehrman S, Ellis MN, Fyfe JA, Otsuka H (1987): Nucleotide sequence changes in thymidine kinase gene of herpes simplex virus type 2 clones from an isolate of a patient treated with acyclovir. Antimicrobial Agent and Chemotherapy 31: 1483-1490.
Klemperer HG, Haynes GR, Shedden WIH, Watson DN (1967): A virus-specific thymidine kinase in BHK-21 cells infected with herpes simplex virus. Virology 34: 120-128.
Larder BA, Darby G (1984): Virus drug-resistance: mechanisms and consequences. Antiviral Research 4: 1-42.
Larder BA, Darby G (1986): Susceptibility to other antiherpes drugs of pathogenic variants of herpes simplex virus selected for resistance to acyclovir. Antimicrobial Agent and Chemotherapy 29: 894-898.
Lindahl T, Adams A, Bjursell G, Bornkamm GW, Kaschka-Dierich C, John U (1976):Covalently closed circular duplex DNA of Epstein-Barr virus in a human lymphoid cell line. Journal of Molecular Biology 102: 511-530.
Littler E, Arrand JR (1988): Characterization of the Epstein-Barr virus-encoded thymidine kinase expressed in heterologous eukaryotic and prokaryotic system. Journal of Virology 62: 3892-3895.
Littler E, Baylis SA, Zeng Y, Conway MJ, Mackett M, Arrand JR (1991): Diagnosis of nasopharyngeal carcinoma by means of recombinant Epstein-Barr virus proteins. Lancet 337: 685-689.
Littler E, Newman W, Arrand JR (1990): Immunological response of nasopharyngeal carcinoma patients to the Epstein-Barr virus-coded thymidine kinase expressed in Escherichia coli. International Journal of Cancer 45: 1028-1032.
Littler E, Zeuthen J, McBride AA, Sorensen ET, Powell KL, Walsh-Arrand JE, Arrand JR (1986): Identification of an Epstein-Barr virus-coded thymidine kinase. EMBO Journal 5: 1959-1966.
Liu MW (1995): Purification and functional studies of the carboxyl terminus of Epstein-Barr virus thymidine kinase . Thesis for Master Degree, Graduate Institute of Microbiology, College of Medicine, National Taiwan University.
Liu MY, Cho SM, Tsai WS, Chang MW, Chen JY, Yang CS (1992): Prevalence of antibody to Epstein-Barr virus DNase in children. Intervirology 34: 192-196.
Liu MY, Pai CY, Shieh SM, Hsu TY, Chen, JY, Yang CS (1992): Cloning and expression of a cDNA encoding the Epstein-Barr virus thymidine kinase gene. Journal of Virological Methods 40: 107-118.
Liu MY, Chang YL, Ma J, Yang HL, Hsu MM, Chen CJ, Chen JY, Yang CS (1997): Evaluation of multiple antibodies to Epstein-Barr virus as markers for detecting patients with nasopharyngeal carcinoma. Journal of Medical Virology 52(3):262-269.
Liu QY, Summers WC (1988): Site-directed mutagenesis of a nucleotide-binding domain in HSV-1 thymidine kinase:effects on catalytic activity. Virology 163: 638-642.
Luka J, Kallin B, Klein G (1979): Induction of Epstein-Barr virus (EBV) cycle in latent infected cells by n-butyrate. Virology 94: 228-231.
Luka J, Siegert W, Klein G (1977): Solubilization of the Epstein-Barr virus-determined nuclear antigen and its characterization as a DNA-binding protein. Journal of Virology 22: 1-8.
Miller RL, Glaser R, Rapp F (1977): Studies of an Epstein-Barr virus induced DNA polymerase. Virology 76: 494-502.
Min KW, Holmquist S, Peiper SC, O’Leary TJ (1991): Poorly differentiated adenocarcinoma with lymphoid stroma (lymphoepithelioma-like carcinomas) of the stomach. Report of three cases with Epstein-Barr virus genome demonstrated by the polymerase chain reaction. American Journal of Pathology 96: 219-227.
Muller CW, Schulz GE (1992): Structure of the complex between adenylate kinase from Escherichia coli and the inhibitor Ap5A refined at 1.9 A resolution: A model for a catalytic transition state. Journal of Molecular Biology 224: 159-177.
Niedobitek G, Young LS, Lau R, Brooks L, Greenspan JS, Rickinson AB (1991): Epstein-Barr virus infection in oral hairy leukoplakia: virus replication in the absence of a detectable latent phase. Journal of General Virology 72: 3035-3046.
Ogino T, Otsuka T, Takahashi M (1977): Induction of deoxypyrimidine kinase activity in human embryonic lung cells infected with varicella-zoster virus. Journal of Virology 21: 1232-1235.
Old JL, Boyse EA, Oettgen HF, De Harven E, Geering G, Willimson B, Clifford P (1966): Precipitating antibody in human antiserum to an antigen present in cultured Burkitt’s lymphoma cells. Proceedings of the National Academy of Sciences USA 56: 1699-1704.
Ooka T, Calender A (1980): Effects of arabinofuranosylthymine on Epstein-Barr virus replication. Virology 104: 219-223.
Pai CY (1992): Cloning and expression of Epstein-Barr virus thymidine kinase gene. Thesis for Master Degree, Graduate Institute of Microbiology, College of Medicine, National Taiwan University.
Raab-Traub N, Hood R, Yang CS, Henry B, Pagano JS (1983): Epstein-Barr virus transcription in nasopharyngeal carcinoma. Journal of Virology 48: 580-590.
Roubal J, Klein G (1981): Synthesis of thymidine kinase(TK) in Epstein-Barr virus-superinfected Raji TK-negative cells. Intervirology 15: 43-48.
Rowe M, Rowe DT, Gregory CD, Young LS, Farrell PJ, Rupani H, Rickinson AB (1987): Difference in B cell growth phenotype reflect novel patterns of Epstein-Barr virus latent gene expression in Burkitt’s lymphoma cells. EMBO Journal 6: 2743-2751.
Sawyer MH, Inchauspe G, Biron KK, Waters DJ, Strans SE, Ostrore JM (1988): Molecular analysis of the pyrimidine deoxyribonucleoside kinase gene of the wild type and acyclovir-resistant strains of varicella-zoster virus. Journal of General Virology 69: 2585-2593.
Sculley TB, Walker PJ, Moss DJ, Pope JH (1984): Identification of multiple Epstein-Barr virus-induced nuclear antigens with sera from patients with rheumatoid arthritis. Journal of Virology 52: 88-93.
Sixbey JW, Nedrud JG, Rabb-Traub N, Hanes RA, Pagano JS(1984): Epstein-Barr virus replication in oropharyngeal epithelial cells. New England Jounral of Medicine 310: 1225-1230.
Sixbey JW, Vesterinen EH, Nedrud JG, Raab-Traub N, Walton LA, Pagano JS(1983): Replication of Epstein-Barr virus in human epithelial cells infected in vitro. Nature(London) 306: 480-483.
Stinchcombe T, Clough W (1985): Epstein-Barr virus induces a unique pyrimidine deoxynucleoside kinase activity in superinfected and virus-producer B cell lines. Biochemistry 24: 2027-2033.
Studier FW (1991): Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system. Journal of Molecular Biology 219: 37-44.
Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW (1990): Use of T7 RNA polymerase to direct expression of cloned genes. Methods in Enzymology 185: 60-89.
Tovey MG, Lenoir G, Begon-Lours J (1978): Activation of latent Epstein-Barr virus by antibody to human IgM. Nature 276: 270-272.
Tsai WS, Chang MW, Chen JY, Lee CY, Liu YG (1989): Seroepidemiological study of Epstein-Barr virus infection in children in Taipei. Acta Paediatrics Sinica 30: 81-86.
Tung PP, Summers WC (1994): Substrate specificity of Epstein-Barr virus thymidine kinase. Antimicrobial Agents and Chemotherapy. 38: 2175-2179.
Veerisetty V, Gentry GA (1985): HSV-1 specific thymidylate kinase activity in infected cells. Intervirology 24: 42-49.
Venables PJW, Roffe LM, Smith PR, Majini RN (1984): Rheumatoid arthritis nuclear antigen and Epstein-Barr virus. Arthritis and Rhumatism 27:476-477.
Wild K, Bohner T, Aubry A, Folkers G, Schulz GE (1995): The three-dimensional structure of thymidine kinase from herpes simplex virus type 1. FEBS Letters 368: 289-292.
Wild K, Bohner T, Folkers G, Schulz GE (1997): The structure of thymidine kinase from herpes simplex virus type 1 in complex with substrates and a substrate analogue. Protein Science 6: 2097-2106.
Wolf H, Seibl R (1984): Benign and malignant disease caused by EBV. Journal of Investigative Dermatology 83 (supp 1.a): 88-95.
Young L, Alfieri C, Hennessy K, Evans H, O’Hara C, Anderson K, Ritz J, Shaprio R, Rickinson A, Kieff R, Cohen J (1989): Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. New England Journal of Medicine 321: 1080-1085.
Zeng Y, Zhong JM, Li LY, Wang PZ, Tang H, Ma YR, Zhu JS, Pan WJ, Liu YX, Wei ZN, Chen JY, Mo YK, Liu EFJ, Tan BF (1983): Follow-up studies of Epstein-Barr virus IgA/VCA antibody-positive persons in Zangwu county, China. Intervirology 20: 190-194.
Zur Hausen H, O’Neill FJ, Freese UK, Hecher E (1978): Presisting oncogenic herpesvirus induced by tumor promoter TPA.Nature 272:373-375.
Zur Hausen H, Schutle-Holthauson H, Klein G, Henle W, Henle G, Clifford P (1970): EBV DNA in biopsies of Burkitt tumors and anaplastic carcinomas of the nasopharynx. Nature 228: 1956-1958.