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研究生:莊馥瑜
研究生(外文):Fu-Yu Chuang
論文名稱:喜姆比蘭嵌紋病毒及木瓜輪點病毒單鏈抗體基因在細菌表現之研究
論文名稱(外文):Studies on expression of scFv antibody gene of Cymbidium mosaic virus and Papaya ringspot virus in bacteria
指導教授:王惠亮
指導教授(外文):Hui-Liang Wang
學位類別:碩士
校院名稱:國立高雄師範大學
系所名稱:生物科學研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:100
中文關鍵詞:喜姆比蘭嵌紋病毒木瓜輪點病毒單鏈抗體重疊延伸的基因裁切法噬菌體表現重組抗體系統
外文關鍵詞:Cymbidium mosaic virusPapaya ringspot virusscFvgene splicing by overlap extensionphage-display system
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喜姆比蘭嵌紋病毒(Cymbidium mosaic virus,CyMV)一直是蘭花栽培者最主要的障礙之一,除了會影響蘭花的生長外,還會造成感染的蘭花品質下降。木瓜輪點病毒(Papaya ringspot virus,PRSV)為木瓜在生長期間最嚴重的病害之一。由喜姆比蘭嵌紋病毒的抗體基因cDNA(CyMV-H 10-1、CyMV-L 23)及木瓜輪點病毒的抗體基因cDNA(PRSV-H 10-9、PRSV-L 3-8)利用聚合酵素反應法(polymerase chain reaction,PCR)分別選殖出重鏈及輕鏈的變異區片段,並藉由重疊延伸的基因裁切法(gene splicing by overlap extension)構築出scFv-CyMV及scFv-PRSV單鏈抗體基因。經核苷酸序列定序分析,得知scFv-CyMV有723個核苷酸,可轉譯成241個胺基酸,而scFv-PRSV有726個核苷酸,可轉譯成242個胺基酸。為了評估scFv-CyMV及scFv-PRSV單鏈抗體蛋白質的表現情形,構築CyMV及PRSV單鏈抗體基因蛋白質表現質體pCANTAB-CyMV及pCANTAB-PRSV,並利用噬菌體表現重組抗體系統分別轉殖至E. coli TG1,並培養出570個及約1242個菌落。由非直接酵素結合抗體檢定法(indirect ELISA)篩選出12個scFv-CyMV(編號C6、C7、C10、C11、C12、C13、C22、C28、C37、C38、C50及C56)及5個scFv-PRSV(P4、P22、P23、P42及P46)呈正反應菌株來進行聚合酶連鎖反應分析及酵素切割。scFv-CyMV及scFv-PRSV聚合酶連鎖反應結果分別為744及747個核苷酸片段,而酵素切割結果兩者皆約1500個核苷酸片段。選取編號為C6、C10、C11的scFv-CyMV轉殖入E. coli HB2151並利用indirect ELISA分析可溶性抗體在細菌的supernatant、periplasmic extract及whole cell extract的表現濃度,結果以位於supernatant最多。為了進行抗體基因轉殖植物和抗體基因蛋白質大量表現,利用聚合酶連鎖反應技術分別完成CyMV和PRSV單鏈抗體基因用來轉殖植物重組質體pBin-19之基因片段,及CyMV和PRSV單鏈抗體基因蛋白質大量表現重組質體pIVEX之基因片段。
Cymbidium mosaic virus(CyMV) is a major limiting factor for orchid plants. Infected plants are unhealthy and produce lower quality flowers. Papaya ringspot virus(PRSV)is a significant pathogen in papaya. The cDNAs of VH and VL of CyMV and PRSV antibody genes were obtained by using polymerase chain reaction. ScFv(single-chain variable fragment)of CyMV and PRSV were constructed by gene splicing by overlap extension (gene SOEing). Results of nucleotide sequences analysis revealed 723 and 726 nucleotides encoding for 241 and 242 amino acids, respectively, for scFv-CyMV and scFv-PRSV. In order to express scFv-CyMV and scFv-PRSV antibody, pCANTAB-CyMV and pCANTAB-PRSV were constructed for transformation into E. cloi TG1 and 570 and 1242 of colonies, respectively, from CyMV and PRSV phage display system were obtained. Twelve scFv-CyMV and five scFv-PRSV colonies were selected with positive reactions in indirect ELISA. 744 and 747 nucleotides fragments of scFv-CyMV and scFv-PRSV, respectively, were cloned from those positively selected colonies by PCR with specific primers. However, two of 1500 nucleotide fragments were obtained for scFv-CyMV and scFv-PRSV, respectively, by using SfiΙand NotΙ enzyme digestion. C6, C10 and C11 of scFv-CyMV were selected for transformation into E. coli HB2151 for monitoring the locations of scFv-CyMV antibody in E. coli cell including supernatant, periplasm and cytosol. The result showed that scFv-CyMV antibody was most aboundant in supernatant. ScFv-CyMV and scFv-PRSV nucleotide fragments were also cloned by PCR with specific primers for inserting into pBin-19 use for transformation of plants and pIVEX-2.4a use for large scale protein expression.
中文摘要……………………………………………………….……..… I
英文摘要………………………………………….………………… III
壹、前言與前人之研究…………………………………………..…….1
貳、材料與方法…………………………………………..………19
一、抗體基因來源…………………………………………19
二、質體DNA的小量抽取……………………………….19
三、單鏈抗體基因之製備………………………….……..20
(一)確定重鏈及輕鏈的變異區位置………………….20
(二)核酸引子之設計………..………………………….21
(三)聚合酶連鎖反應………………………………..….22
(四)聚合酶連鎖反應產物之電泳膠體分析..………………27
(五)DNA片段的回收與純化….…………………………...27
(六)重疊延伸的基因裁切法……………………………….28
四、單鏈抗體核酸序列之譯讀………………………….…….29
五、單鏈抗體核酸序列之分析………………………….……….32
六、表現單鏈抗體基因蛋白質質體之構築………….……….32
(一)酵素切割………….………………………………..32
(二)黏接反應…………………………………………...33
七、轉殖反應…………………………….………….……….33
(一)勝任細胞………….………………………………..33
(二)轉殖反應…………………………………………...36
八、重組嗜菌體之製備………………….………….……….36
九、篩選反應…….………………….………….……………37
(一)以PEG沉澱重組噬菌體………………………………..37
(二)固體相的篩選…..………………………..………..37
(三)以重組噬菌體再次感染E. coli………………………..39
十、利用ELISA篩選出可與抗原有反應的菌株....……………39
(一)準備可溶性抗體…...………………………………..39
(二)進行ELISA篩選步驟…………………..……………..40
十一、準備重組噬菌體感染E. coli HB2151....……………41
十二、可溶性抗體製備……………………......……………41
(一)製備於supernatant的可溶性抗體…………………….41
(二)製備於perplasmic extract的可溶性抗體……………..42
(三)製備於whole cell extract的可溶性抗體…………….42
十三、確定單鏈抗體蛋白質的表現………......……………42
(一)聚合酶連鎖反應………………………………..….42
(二)酵素切割………….………………………………..43
(三)酵素結合抗體檢定法.……………………………….43
十四、重組質體之構築…………..……......……………43
(一)構築表現抗體基因重組質體………………....43
(二)構築抗體基因轉殖植物重組質體………………....47
(三)黏接反應…………………………………………...50
參、結果………………………………………………………….…51
一、聚合酶連鎖反應產物及gene SOEing結果之電泳分析…..51
二、單鏈抗體基因之定序與譯讀……………..……………..53
三、篩選後所得到的結果……………..……………………..59
四、以酵素結合抗體檢定法分析單鏈抗體的表現..………..59
五、確定單鏈抗體蛋白質的表現………......………………61
(一)聚合酶連鎖反應………………………………..….61
(二)酵素切割………….………………………………..67
(三)酵素結合抗體檢定法.……………………………….70
六、重組質體之構築…………..……......……………….…70
肆、討論…………………………………………….…………………76
伍、參考文獻……………………………………………………………81
陸、附錄…………………………………………………………………93
王惠亮 王嬿婷 廖麗貞。2000。核酸探針雜合反應或反轉錄聚合酶連鎖反應偵測喜姆比蘭嵌紋病毒之比較。植物保護學會會刊 42:25-42。
王嬿婷。1996。蘭花喜姆比蘭嵌文病毒偵測技術之比較與應用。國立高雄師範大學科學教育研究所碩士論文。
包慧俊。2000。木瓜輪點病毒鞘蛋白轉基因木瓜抗病性狀之研究。國立中興大學植物病理研究所博士論文。
李惠鈴。1990。蝴蝶蘭病毒之特性及其防治。國立中興大學植物病理研究所碩士論文。
李慶孝。1999。喜姆比蘭嵌紋病毒及木瓜輪點病毒抗體基因在細菌之表現和木瓜輪點病毒抗體基因在木瓜之轉殖研究。國立高雄師範大學生物科學研究所碩士論文。
位國慶。1995。台灣蝴蝶蘭病毒之發生與防治。農業世界 137:84-88。
張清安。1996。蘭花病毒病之特性及其防治。農業世界 137:14-20。
葉錫東。1999。轉基因作物應用於植物保護之現況。植物保護學會會刊。 41:87-106。
賴本智。1996。台灣蘭花產業的過去、現在與未來。農業世界 137:7-10。
Aaziz, R., and Tepfer, M. 1999. Recombination in RNA viruses and in virus-resistant transgenic plants. J. Gen. Virol. 80:1339-1346.
Abel, P. P., Nelson, R. S., De, B., Hoffmann, N., Rogers, S. G., Fraley, R. T., and Beachy, R. N. 1986. Delay of disease development in transgenic plant that express the tobacco mosaic virus coat protein gene. Science 232: 738-743.
Alfthan, K., Takkinen, K., Sizmann, D., Soderlund, H., and Teeri, T. T. 1995. Properties of a single-chain antibody containing different linker peptides. Protein Eng. 8:725-731.
Anamthawat, K., and Vajrabhaya, T. 1979. Survey of the occurrence of cymbidium mosaic virus in dendrobium orchids in Bankok. Aust. Orchid Rev. 44:79-81.
Anderson, J. M., Palukaitis P., and Zaitlin M. 1992. A defective replicase gene induces resistance to cucumber mosaic virus in transgenic tobacco plant. Proc. Natl. Acad. Sci. U.S.A. 89:8759-8763.
Argos, P. 1990. An investigation of oligopeptides linking domains in protein tertiary structures and possible candidates for general gene fusion. J. Mol. Biol. 211:943-958.
Batra, J. K., Kasprzyk, P. G., Bird, R. E., Pastan, I., and King, R. C. 1992. Recombinant anti-erbB2 immunotoxins containing Pseudomonas exotoxin. Proc. Natl. Acad. Sci. U.S.A. 89:5867-5871.
Baulcombe, D. 1994. Replicase mediated resistance: a novel type of virus resistance in transgenic plant. Trends Microbiol. 2:60-63.
Baulcombe, D. C. 1996. RNA as a target and an initiator of posttranscriptional gene silencing in transgenic plants. Plant Mol. Biol. 32:79-88.
Beachy, R. N. 1990. Coat protein mediated resistance against virus infection. Ann. Rev. Phytopathol. 28:451-474.
Benvenuto, E., and Tavladoraki, P. 1995. Immunotherapy of plant viral diseases. Trends Microbiol. 3:272-275.
Benvenuto, E., Ordas, R. J., Tavazza, R., Ancora, G., Biocca S., Cattaneo, A., and Galeffi, P. 1991. Phytoantibodies:A general vector for the expression of immunoglobulin domains in transgenic plants. Plant Mol. Biol. 17:865-874.
Bird, R.E., Hardman, K. D., Jacobson, J. W., Johnson, S., Kaufman, B. M., Lee, S. M., Lee, T., Pope, S.H., Riordan, G. S., and Whitlow, M. 1988. Single-chain antigen-binding proteins. Science 21:423-426.
Boss, A., Kenten, J. H., Wood, C. R., and Emtage, J. S. 1984. Assembly of functional antibodies from immunoglobulin heavy and light chains synthesized in E. coli. Nucleic Acids Res. 12:3791-3806.
Brinkmann, U., Reiter, Y., Jung, S. H., Lee, B., and Pastan, I. 1993. A recombinant immunotoxin containing a disulfide-stabilized Fv fragment. Proc. Natl. Acad. Sci. U.S.A. 90:7538-7542.
Buchner, J., Pastan, I., and Brinkmann, U. 1992. A method to increase the yield of properly folded recombinant fusion proteins:Single-chain immunotoxins from renaturation of bacterial inclusion bodies. Anal. Biochem. 205:263-270.
Cabilly, S., Riggs, A. D., Pande, H., Shively, J. E., Holmes, W. E., Rey, M. Perry, L. J., Wetzel, R. and Heyneker H. L. 1984. Generation of antibody activity from immunoglobulin polypeptide chains produced in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 81:3273-3277.
Carr, J. P., and Zaitlin, M. 1994. Replicase mediated resistance. Semin. Virol. 4:339-347.
Chia, T. F., and Chan, Y. S. 1992. Chracterization of cymbidium mosaic virus coat protein gene and its expression in transgenic tobacco plant. Plant Mol. Biol. 18:1091-1099.
Colcher, D., Milenic, D. E., Ferroni, P., Carrasquillo, J. A., Reynolds, J. C., Roselli, M., Larson, S. M., and Schlom, J. 1990. In vivo fate of monoclonal antibody B72.3 in patients with colorectal cancer. J. Nucl. Med. 31:1133-1142.
Compbell, N. A. 1993. Biology. Ch17. 3rd ed. California. The Benjamin/ Cummings Publishing Company, Inc.
Cooper, B., Lapidot, M., Herick, J. A., Dodds J. A., and Beachy, R. N. 1995. A defective movement protein of TMV in transgenic plants confers resistance to multiple viruses whereas the functional analog increases susceptibility. Virology 206:307-313.
De Jaeger, G., Buys, E., Eeckhout, D., Bruyns, A. M., De Neve, M., De Wilde, C., Gerats, T., Van Montagu, M., Fischer, R., and Depicker, A. 1997. Use of phage display for isolation and characterization of single-chain variable fragments against dihydroflavonol 4-reductase from Petunia hybrida. FEBS Lett. 403:116-122.
De Jaeger, G., De Wilde, C., Eeckhout, D., Fiers, E., and Depicker, A. 2000. The plantibody approach: expression of antibody genes in plants to modulate plant metabolism or to obtain pathogen resistance. Plant Mol. Biol. 43:419- 428.
De Neve, M., De Loose, M., Jacobs, A., Van Houdt, H., Kaluza, B., Weidle, U., Van Montagu, M., and Depicker, A. 1993. Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis. Transgenic Res. 2:227-237.
Desplancq, D., King, D. J., Lawson, A. D., and Mountain, A. 1994. Multimerization behaviour of single chain Fv variants for the tumour-binding antibody B72.3. Protein Eng. 7:1027-1033.
Duenas, M., and Borrebaeck, C. A. K. 1994. Clonal selection and amplification of phage displayed antibodies by linking antigen recognition and phage replication. Bio/Technology 12:999-1002.
During, K. Hippe, S., Kreuzaler, F., and Schell, J. 1990. Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Plant Mol. Biol. 15:281-293.
Edelman, G. M. 1973. Antibody structure and molecular immunology. Science 180:830-840.
Faccioli, G., and Marani, F. 1979. Cymbidium mosaic virus associated with flower necrosis in Cattleya orchids. Phytopath. Medit. 18:21-25.
Firek, S., Draper, J., Owen, M. R., Gandecha, A., Cockburn, B., and Whitelam, G. C. 1993. Secretion of a functional single-chain Fv protein in transgenic tobacco plants and cell suspension cultures. Plant Mol. Biol. 23:861-870.
Fitch, M. M. M., Manshardt, R. M., Gonsalves, D., Slightom, J. L., and Sanford, J. C. 1992. Virus resistant papaya plants derived from tissues bombarded with the coat protein gene of papaya ringspot virus. Bio/Technology 10:1466-1472.
Fitchen, J. H., and Beachy, R. N. 1993. Genetically engineered protection against viruses in transgenic plants. Annu. Rev. Microbiol. 47:739-763.
Frowd, J. A., and Tremaine, J. H. 1977. Physical, chemical, and serological properties of cymbidium mosaic virus. Phytopathology 67:43-49.
Glockshuber, R., Malia, M., Pfitzinger, I., and Pluckthun, A. 1990. A comparison of strategies to stabilize immunoglobulin Fv-fragments. Biochemistry 29:1362-1367.
Gonsalves, D., and Slighton, J. L. 1993. Coat protein — mediated protection:Analysis of transgenic plants for resistance in a variety of crops. Semin. Virol. 4:397-405.
Hari, V., Siegel , A., Rozek, C., and Timerlake, W. E. 1979. The RNA of tobacoo etch virus contain poly(A). Virology 92:568-571.
Hiatt, A., Cafferkey, R., and Bowdish, K. 1989. Production of antibodies in transgenic plants. Nature 342:76-78.
Hoogenboom, H. R. 1997. Designing and optimizing library selection strategies for generating high-affinity antibodies. Trends Biotechnol. 15:62-70.
Horton, R. M., Ho, S. N., Pullen, J. K., Hunt, H. D., Cai, Z., and Pease, L. R. 1993. Gene splicing by overlap extension. Methods Enzymol. 217:270-279.
Horton, R. M., Hunt, H. D., Ho, S. N., Pullen, J. K., and Pease, L. R. 1989. Engineering hybrid genes without the use of restriction enzymes:gene splicing by overlap extension. Gene 77:61-68.
Huston, J. S., Levinson, D., Mudgett-Hunter, M., Tai, M. S., Novotny, J., Margolies, M. N., Ridge, R. J., Bruccoleri, R. E., Haber, E., and Crea, R. 1988. Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 85:5879-5883.
Ishii, M., and Holtzmann, O. V. 1963. Papaya mosaic disease in Hawaii. Plant Dis. Rep. 49:947951.
Jan, F. J., Pang, S. Z., Fagoaga, C., and Gonsalves D. 1999. Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene silencing. Transgenic Res. 8:203-213.
Jensen, D. D. 1949. Papaya ringspot virus and its insect vector relationships. Phytopathology 39:212-220.
Jensen, D. D. 1951. Mosaic or black streak disease of cymbidium orchids. Phytopathology 41:401-414.
Kawano, S., and Yonaha, T. 1992. The occurrence of papaya leaf-distortion mosaic virus in Okinawa. Tech. Bull. of FFTC 132:13-23.
Lapidot, M., Gafny, R., Ding, B., Wolf, S., Lucas, W. J., and Beachy, R. N. 1993. A dysfunctional movement protein of tobacco mosaic virus that partially modifies the plasmodesmata and limits virus spread in transgenic plant. Plant J. 4:959-970.
Lawson, R. H., and Hearon, S. S. 1973. Symptomatology of cattleya mericlones infected with cymbidium mosaic virus. Amer. Orchid Soc. Bull. 42:1071-1074.
Lin, C. C. 1980. Strains of papaya ringspot virus and their cross protection. Ph. D. thesis. National Taiwan University, Taipei.
Lindbo, J. A., and Dougherty, W. G. 1992a. Pathgen-derived resistance to a potyvirus:immune and resistant phenotypes in transgenic tobacco expressing altered forms of a potyvirus coat protein nucleotide sequence. Mol. Plant Microb. Interact. 5:144-153.
Lindbo, J. A., and Dougherty, W. G. 1992b. Untranslatable transcripts of the tobacco etch virus coat protein gene sequence can interfere with tobacco etch virus replication in transgenic plants and protoplasts. Virology 189:725-733.
Lomonossoff, G. P. 1995. Pathgen-derived resistance to plant viruses. Annu. Rev. Phytopathol. 33:323-343.
McCafferty, J., Griffiths, A. D., Winter, G., and Chiswell, D. J. 1990. Phage antibodies:filamentous phage displaying antibody variable domains. Nature 348:552-554.
Messing, J. 1991. Cloning in M13 phage or how to use biology at its best. Gene 100:3-12.
Neo, K. K., Wong, S. M., and Wu, M. 1993. Nucleotide sequence and in vitro translation of the coat protein gene of cymbidium mosaic virus. Virus Genes. 7:157-170.
Okemura, A. K., Kamemoto, H., and Ishii, M. 1984. Incidence and expression of cymbidium mosaic virus in Dendrobium hybrid. HITAHR, College of tropical agriculture and human resources. University of Hawii.
Owen, M., Gandecha, A., Cockburn, B., and Whitelam, G. 1992. Synthesis of a function anti-phytochrome single-chain Fv protein in transgenic tobacco. Bio/Technology 10:790-794.
Pang, S Z., Jan, F. J., and Gonsalves, D. 1997. Nontarget DNA sequences reduce the transgene length necessary for RNA-mediated tospovirus resistance in transgenic plants. Proc. Natl. Acad. Sci. U.S.A. 94:8261-8266.
Pisi, A., Marani, F., and Bertaccini, A. 1982. Intracellular inclusions in host plants infected with cymbidium mosaic virus. Phytopath. Medit. 21:27-30.
Powell, P. A., Stark, D. M., Sanders, P. R., and Beachy R. N. 1989. Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA. Proc. Natl. Acad. Sci. U.S.A. 86:6949-6952.
Purcifull, D. E., Edwardson, J. R., Hiebert, E., and Gonsalves, D. 1984. Papaya ringspot virus. CMI/AAB Descriptions of plant Viruses. No.84
Rasched, I., and Oberer, E. 1986. Ff coliphages: structural and functional relationships. Microbiol. Rev. 50:401-427.
Reiter, B., Brinkmann, U., Jung, S. H., Lee, B., Kasprzyk, P. G., King, R. C. and Pastan, I. 1994a. Improved binding and anti tumor activity of a recombinant anti-erbB2 immunotoxin by disulfide-stabilization of the Fv fragment. J. Biol. Chem. 269:18327-18331.
Reiter, B., Brinkmann, U., Jung, S. H., Lee, B., and Pastan, I. 1994b. Engineering disulfide bonds into conserved framework regions of Fv fragments:Recombinant immunotoxins containing disulfide-stabilized Fv with improved biochemical characteristics. Protein Eng. 7:697-704.
Riechmann, J. L., Lain, S., and Garcia, J. A. 1992. Highlights and prospects of potyvirus molecular biology. J. Gen. Virol. 73:1-16.
Riechmann, L., Foote, J., and Winter, G. 1988. Expression of an antibody Fv fragment in myeloma cells. J. Mol. Biol. 203:825-828.
Robinson, D. J. 1996. Environmental risk assessment of releases of transgenic plants containing virus-derived inserts. Transgenic Res. 5:359-562.
Sanford, J. C., and Johnston, S. A. 1985. The concept of parasite — derivated resistance: deriving resistance genes from the parasite''s own genome. J. Thero. Biol. 113:395-405.
Sanger, F., Nicklen, S., and Coulson, A. R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. 74:5463-5467.
Sastry, L., Alting-Mees, M., Huse, W. D., Short, J. M., Sorge, J. A. , Hay, B. N., Janda, K. D., Benkovic, S. J., and Lerner R. A. 1989. Cloning of the immunogical repertoire in Escherichia coli for generation of monoclonal catylytic antibodies: Construction of a heavy chain variable region-specific cDNA library. Proc. Natl. Acad. Sci. U.S.A. 86:5728-5732.
Singh, A. B. 1971. Transmission of papaya leaf reduction virus by Myzus persicae. Plant Dis. Rep. 55:526-529.
Skerra, A., and Pluckthun, A. 1988. Assembly of a functional immunoglobulin Fv fragment in E. coli. Science 240:1038-1041.
Tavladoraki, P., Benvenuto, E., Trinca, S., De Martinis, D., Cattaneo, A., Galeffi, P. 1993. Transgenic plants expressing a functional single-chain Fv antibody are specifically protected from virus attack. Nature 366:469-472.
Tepfer, M. 1993. Viral genes and transgenic plants: what are the potential environmental risk? Bio/Technology 11:1125-1132.
Tyutyulkova, S., and Paul., S. 1994. Selection of functional human immunoglobulin light chains from a phage-display library. Appl. Biochem. Biotechnol. 47:191-198.
van den Boogaart, T., Lomonossoff, G. P., and Davies, J. W. 1998. Can we explain RNA-mediated virus resistance by homology-dependent gene silencing? Mol. Plant Microbe. Interact. 11:717-723.
Vaucheret, H., Christophe, B., Elmayan, T., Feuerbach, F., Godon, C., Morel, J. B., Mourrain, P., Palauqui, J. C., and Vernhettes, S. 1998. Transgene-induced gene silencing in plants. Plant J. 16:651-659.
Wang, H. L., Wang C. C., Chiu, R. J., and Sun, M. H. 1978. Preliminary study on papaya ringspot virus in Taiwan. Plant Prot. Bull. 20:133-140.
Wang, H. L., Yeh, S. D., Chiu, R. J., and Gonsalves, D. 1987. Effectivess of cross-protection by mild mutants of papaya ringspot virus for control of ringspot disease of papaya in Taiwan. Plant Dis. 71:491-497.
Whitlow, M., Bell, B. A., Feng, S. L., Filpula, D., Hardman, K. D., Hubert, S. L., Rollence, M.L., Wood, J. F., Schott, M. E., and Milenic, D. E. 1993. An improved linker for single-chain Fv with reduced aggregation and enhanced proteolytic stability. Protein Eng. 6:989-995.
Whitlow, M., Filpula, D., Rollence, M. L., Feng, S. L., and Wood, J. F. 1994. Multivalent Fvs: characterization of single-chain Fv oligomers and preparation of a bispecific Fv. Protein Eng. 7:1017-1026.
Wood, C. R., Boss, M. A., Kenten, J. H., Calvert, J.E., Roberts, N. A., and Emtage, J. S. 1985. The synthesis and in vivo assembly of functional antibodies in yeast. Nature 314:446-449.
Yeh, S. D., and Gonsalves, D. 1984. Evaluation of induced mutants of papaya ringspot virus for control by cross protection. Phytopathology 74:1086-1091.
Yeh, S. D., and Gonsalves, D. 1994. Practies and perspective of control of papaya ringspot virus by cross protection. Adv. Dis. Vector Res. 10:237-257.
Yeh, S. D., Gonsalves, D., Wang, H. L., Namba, R., and Chiu, R. J. 1988. Control of papaya ringspot virus by cross protection. Plant Dis. 72. 375-380.
Yokota, T., Milenic, D. E., Whitlow, M., and Schlom, J. 1992. Rapid tumor penetration of a single-chain Fv and comparison with other immunoglobulin forms. Cancer Res. 52:3402-3408.
Yuen, C. K. K. H., Kamemoto, H., and Ishii, M. 1979. Transmission of cymbidium mosaic virus through seed propagation in Dendrobium. Amer. Orchid Soc. Bull. 48:1245-1248.
Zettler, F. W., Ko, N. J., Wisler, G. C., Elliott, M. S., and Wong, S. M. 1990. Viruses of orchids and their control. Plant Dis. 74:621-626.
Zhou, H., Fisher, R. J., and Papas, T. S. 1994. Optimization of primer sequences for mouse scFv repertoire display library construction. Nucleic Acids Res. 22:888-889.
Zimmermann, S., Schillberg, S., Liao, Y.C., and Fisher, R. 1998. Intracellular expression of TMV-specific single-chain Fv fragments leads to improved virus resistance in Nicotiana tabacum. Molecular Breeding 4:369-379.
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