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研究生:唐為瑄
研究生(外文):Wei-Hsuan Tang
論文名稱:以哺乳類細胞產製長效型豬干擾素α基因重組蛋白及其特性分析
論文名稱(外文):Cloning, Expression, Purification and Antiviral Activity Assesment of Recombinant Porcine IFNα-IgG Fusion Protein Expressed by Mammalian Cells
指導教授:郭村勇
指導教授(外文):Tsun-Yung Kuo
口試委員:郭村勇賴秀穗鄭金益
口試委員(外文):Tsun-Yung KuoShiow-Suey LaiJin-Yi Cheng
口試日期:2011-07-25
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:生物技術研究所碩士班
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:73
中文關鍵詞:干擾素豬生殖與呼吸道綜合症病毒
外文關鍵詞:interferon, Porcine reproductive and respiratory syndrome virus
相關次數:
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以哺乳類細胞表現基因重組蛋白可正確折疊並進行轉譯後修飾作用,對蛋白
質的生物活性有很大的影響性。本論文選用中國倉鼠卵巢細胞株(CHO cell)表現
長效型豬干擾素α 重組蛋白。
干擾素的半衰期短,因此本實驗將豬干擾素α 基因與豬IgG Fc 片段基因進
行連接,構築長效型的豬干擾素α 基因,經序列比對確認無誤後,轉染至中國倉
鼠卵巢細胞株(CHO cell)中,表現出豬長效型干擾素α 基因重組蛋白。經酵素連
結免疫吸附分析法(Enzyme-Linked Immunosorbent Assay ELISA)確認為分泌型 P
IFNα-Fc 基因重組蛋白,該分泌蛋白可分別以鎳金屬親和性管柱或 Protein G 親
合性管柱純化出。
以豬生殖與呼吸道綜合症(Porcine reproductive and respiratory syndrome,
PRRS)病毒感染Marc145細胞模式分析長效型豬干擾素α 重組蛋白之抗病毒活
性,發現 49 nM 就對該病毒有抑制效果。分別以 175 平方公分角瓶和轉瓶培養
產製長效型豬干擾素α 重組蛋白,結果角瓶培養可產製出約 450 μg/ml 以上的長
效型豬干擾素α 重組蛋白,而轉瓶培養可產製出約 800 μg/ml 的長效型豬干擾素
α 重組蛋白。抗病毒活性測試部份,無論是以 175 平方公分角瓶或轉瓶培養所
產製的長效型豬干擾素α,生產長效型豬干擾素α時,細胞所分泌的干擾素抗病
毒活性皆會隨著細胞培養時間的增長而增高直到細胞脫落死亡為止,其抗 PRRS
病毒活性分別為 691 U ml-1 和 2430 U ml-1,轉瓶培養方式所產製出的長效型豬
干擾素α 的抗病毒活性明顯高於以 175 平方公分角瓶培養。
Mammalian cell expression system can yield properly-folded and
post-translationally modified recombinant proteins which benefits greatly the
biological activity of proteins. In this study, Chinese hamster ovary (CHO) cells were
selected for the expression of recombinant interferon-α (IFNα) proteins with extended
serum half-life.
Due to the short serum half-life of IFNα, a glycine-serine linker was used to fuse
genes of porcine IFNα and the Fc region of porcine IgG antibody for the construction
of IFNα with extended serum half-life. The genes were cloned, fused, confirmed with
sequence analysis, and subsequently transfected to CHO cells. By ELISA evaluation,
PIFNα-Fc recombinant proteins were confirmed as secreted proteins. Secreted proteins
were then purified using nickel and Protein G affinity columns.
Porcine reproductive and respiratory syndrome virus (PRRSV) was used in
assaying the antiviral activity of IFNα with extended half-life. In measuring interferon
activity, results showed that 49 nM of IFNα with extended half-life sufficiently
displayed antiviral effects against PRRSV. For recombinant protein expression and
production of IFNα with extended half-life, 175 cm2 flasks and roller bottles were used.
175 cm2 flasks and roller bottles produced more than 450 μg/ml and about 800 μg/ml
of IFNα with extended half-life recombinant proteins, respectively. In the antiviral
assay, antiviral effects of IFNα with extended half-life produced in either 175 cm2
flasks or roller bottles were shown to increase over time with longer periods of cell
culturing. Furthermore, the antiviral activities of the interferons produced were found
to be 691 U ml-1 and 2430 U ml-1, respectively. The antiviral activities of recombinant
IFNα proteins with extended half-life produced in roller bottles were significantly
higher than those produced in 175 cm2 flasks.
摘要........……………………………………………………………………………...i
Abstract……………………………………………………………………………….ii
目錄(Table of contents)…………………………………………………...………….iii
表目錄(List of table)………………………………………………………..………..vii
圖目錄(List of Figures)……………………………………………………..….……viii
壹、緒言………………………………………………………………...…….………..1
貳、文獻探討……………………………………………………………….….……….3
一、干擾素的簡介………………………………………………………….…….….…3
1.1 干擾素之發現……………………………………………………….….………..3
1.2 干擾素之分類……………………………………………………….…..………..3
二、干擾素之誘發機製………………………………………………….…….……….3
三、干擾素抗病毒機製探討………………………………………………..…….……4
3.1 干擾素誘發抗病毒蛋白之訊息傳導路徑…………………………….…………4
3.2 抗病毒蛋白抗病毒機制……………………………………………….……….…..5
四、豬生殖與呼吸綜合症病毒之介紹…………………………………..…....……..6
五、哺乳動物細胞表現系統…………………………….……………….……….….7
六、干擾素的半衰期...........................................................8
6.1 免疫球蛋白 IgG Fc 可延長生物分子之半衰期.....................................8
6.2 IgG 的接受體 FcRn 是使具 Fc 片段生物分子半衰期延長之因素..................9
参、材料與方法..............................................................11
第一節 長效型豬干擾素α基因之次選殖...............................................11
一、 長效型豬干擾素α 之模板DNA 製備............................................11
二、 特異性引子的設計.....................................................11
三、 以PCR 法增幅長效型豬干擾素α基因片段......................................12
四、 PCR 產物的確認…………………………………………..……………...12
五、 PCR 產物的純化(膠體萃取法)………………………………….……….....12
六、 限制酵素切割反應………………………………………………….…........13
七、 接合反應………………………………………………………………...….....13
八、 勝任細胞的製備.......................................................13
九、 細菌轉形作用……………………………………………………………….14
十、 重組質體的挑選及確認…………………………………………………........14
十一、 序列分析………………………………………………………………….14
第二節 長效型豬干擾素α基因重組蛋白之表現…………………………………15
一、 以鹼溶解法萃取長效型豬干擾素α之質體DNA………………………...…..15
二、 長效型豬干擾素α 基因轉染至中國倉鼠卵巢細胞株…………………..…15
三、 以Zeocin 抗生素篩選長效型豬干擾素α 重組基因的CHO 細胞株………16
四、 間接免疫螢光染色確認長效型豬干擾素α 重組蛋白表現….......................16
五、 以酵素連結免疫分析(ELISA)法檢測長效型豬干擾素α 重組蛋白表現.…17
六、 十二烷基硫酸鈉-聚丙烯醯胺膠電泳分析和西方墨點轉漬法分析確認長效
型豬干擾素α基因重組蛋白的表現………………..……………………….17
6.1 十二烷基硫酸鈉-聚丙烯醯胺膠電泳分析 (SDS-PAGE)……………………17
6.2 西方墨點法確認重組蛋白表現……………………………………………….18
第三節 長效型豬干擾素α基因重組蛋白之純化………………………………..19
一、 以三明治 ELISA 法測定長效型豬干擾素α產製細胞的生長和重組蛋白表
現量的關係………………………………………………………………….19
二、 長效型豬干擾素α 基因重組蛋白的純化……………………………………20
2.1 以 Protein G column 純化之 P IFNα-Fc 基因重組蛋白……………………...20
2.2 以His column純化之IFN α Linker IgG基因重組蛋白…………………….….20
2.3 純化後重組蛋白質透析……………………………………………………….20
三、蛋白質定量………………………………………………………………………21
第四節 長效型豬干擾素α 的抗 PRRSV 活性測試……………………….…..21
一、細胞及病毒…………………………………………………………………..…..21
1.1 MARC-145 細胞株培養…………………………………………………...…..21
二、長效型豬干擾素α 抗病毒效果測試………………………………….……...…22
第五節 以角瓶及轉瓶產製長效型豬干擾素α……………………………….….22
一、以175平方公分角瓶細胞培養方式產製長效型豬干擾素α基因重組蛋白…...22
二、以轉瓶培養方式來產製長效型豬干擾素α基因重組蛋白……………….…….23
三、以三明治 ELISA 法測定長效型豬干擾素α 之表現量……………………….24
四、長效型豬干擾素α基因重組蛋白之活性測定……………………..……………24
4.1干擾素活性計算方法………………………….………………………………..25
肆、結果…………………………………………………………………………..….27
第一節 長效型豬干擾素α基因之次選殖……………………...………….……....27
第二節 長效型豬干擾素α基因重組蛋白之表現………….………………......….27
一、 以間接免疫螢光染色法確認長效型豬干擾素α基因重組蛋白之表現…….27
二、 長效型豬干擾素α重組基因細胞之篩選……………………….……...…...28
三、 以酵素連結免疫分析(ELISA)檢測長效型豬干擾素α基因重組蛋白表現....28
第三節 長效型豬干擾素α 基因重組蛋白之初步純化與鑑定……………….…..28
一、以三明治 ELISA 法測定不同培養時段長效型豬干擾素α 基因的表現情形28
二、以 protein G 和鎳金屬親和性管柱進行長效型豬干擾素純化……………....29
第四節 長效型豬干擾素α 的抗 PRRSV 活性測試………………………...…29
第五節 以角瓶及轉瓶產製長效型豬干擾素α之比較…………………………….30
一、 長效型豬干擾素α 重組蛋白產製之曲線圖…………………………………30
二、 以 175 平方公分角瓶和轉瓶培養方式產製長效型豬干擾素α 基因重組蛋
白之活性測定……………………………………………………………….30
伍、討論………………………………………………………………………………31
附錄……..…………………………………………………………………………..39
參考文獻 ………………………………………………………………..………...…57

吳忠晉。豬干擾素α基因重組蛋白誘發細胞抗PRRSV效力之探討。國立宜蘭大學
生物技術研究所碩士論文(2007)。
陳漢婷。 豬干擾素α基因重組蛋白的產製、純化和活性分析。國立宜蘭大學生物
技術研究所碩士論文(2008)。
Ahouse, J.J. et al. Mouse MHC class I-like Fc receptor encoded outside the MHC. J
Immunol 151, 6076-6088 (1993).
Aixinjueluo, W. et al. Mechanisms for the apoptosis of small cell lung cancer cells
induced by anti-GD2 monoclonal antibodies: roles of anoikis. J Biol Chem 280,
29828-29836 (2005).
Akilesh, S. et al. The MHC class I-like Fc receptor promotes humorally mediated
autoimmune disease. J Clin Invest 113, 1328-1333 (2004).
Akira, S. Mammalian Toll-like receptors. Curr Opin Immunol 15, 5-11 (2003).
Albina, E., Carrat, C. & Charley, B. Interferon-alpha response to swine arterivirus
(PoAV), the porcine reproductive and respiratory syndrome virus. J Interferon
Cytokine Res 18, 485-490 (1998).
Andreakos, E.T., Foxwell, B.M., Brennan, F.M., Maini, R.N. & Feldmann, M.
Cytokines and anti-cytokine biologicals in autoimmunity: present and future. Cytokine
Growth Factor Rev 13, 299-313 (2002).
Asker, N., Baeckstrom, D., Axelsson, M.A., Carlstedt, I. & Hansson, G.C. The human
MUC2 mucin apoprotein appears to dimerize before O-glycosylation and shares
epitopes with the 'insoluble' mucin of rat small intestine. Biochem J 308 ( Pt 3),
873-880 (1995).
Bautista, E.M. & Molitor, T.W. IFN gamma inhibits porcine reproductive and
respiratory syndrome virus replication in macrophages. Arch Virol 144, 1191-1200
(1999).
Berenguer, J. et al. Pegylated interferon {alpha}2a plus ribavirin versus pegylated
interferon {alpha}2b plus ribavirin for the treatment of chronic hepatitis C in
HIV-infected patients. J Antimicrob Chemother 63, 1256-1263 (2009).
Bergtold, A., Desai, D.D., Gavhane, A. & Clynes, R. Cell surface recycling of
internalized antigen permits dendritic cell priming of B cells. Immunity 23, 503-514
(2005).
Bialy H. Recombinant proteins: Virtual authenticity. Bio/Technology 5:883–890.
(1987).
Billiau, A., Heremans, H., Allens, P.T., Baron, S. & de Somer, P. Interferon inhibits
C-type virus at a posttranscriptional, prerelease step. Arch Virol 57, 205-220 (1978).
Billiau, A. Interferon: the pathways of discovery I. Molecular and cellular aspects.
Cytokine Growth Factor Rev 17, 381-409 (2006).
Buckholz, R.G. Yeast systems for the expression of heterologous gene products. Curr
Opin Biotechnol 4, 538-542 (1993).
Buckwold, V.E. et al. Safety pharmacology, toxicology and pharmacokinetic
assessment of recombinant human omega-interferon produced from CHO-SS cells.
Basic Clin Pharmacol Toxicol 99, 62-70 (2006).
Buckwold, V.E. et al. Antiviral activity of CHO-SS cell-derived human omega
interferon and other human interferons against HCV RNA replicons and related
viruses. Antiviral Res 73, 118-125 (2007).
Burmeister, W.P., Huber, A.H. & Bjorkman, P.J. Crystal structure of the complex of
rat neonatal Fc receptor with Fc. Nature 372, 379-383 (1994).
Cafruny, W.A. et al. Porcine reproductive and respiratory syndrome virus (PRRSV)
infection spreads by cell-to-cell transfer in cultured MARC-145 cells, is dependent on
an intact cytoskeleton, and is suppressed by drug-targeting of cell permissiveness to
virus infection. Virol J 3, 90 (2006).
Cantell, K., Hirvonen, S., Kauppinen, H.L. & Myllyla, G. Production of interferon in
human leukocytes from normal donors with the use of Sendai virus. Methods Enzymol
78, 29-38 (1981).
Carpick, B.W. et al. Characterization of the solution complex between the
interferon-induced, double-stranded RNA-activated protein kinase and HIV-I
trans-activating region RNA. J Biol Chem 272, 9510-9516 (1997).
Cavanagh, D. Nidovirales: a new order comprising Coronaviridae and Arteriviridae.
Arch Virol 142, 629-633 (1997).
Chang, T. W. and Yu, L., Hybrid with interferon-alpha and an immunoglobulin Fc
linked through a non-immunogenic peptide. United state patent 5723125 (1998).
Chaudhury, C. et al. The major histocompatibility complex-related Fc receptor for IgG
(FcRn) binds albumin and prolongs its lifespan. J Exp Med 197, 315-322 (2003).
Cheon, D.S., Chae, C. & Lee, Y.S. Detection of nucleic acids of porcine reproductive
and respiratory syndrome virus in the lungs of naturally infected piglets as determined
by in-situ hybridization. J Comp Pathol 117, 157-163 (1997).
Chinsangaram, J., Piccone, M.E. & Grubman, M.J. Ability of foot-and-mouth disease
virus to form plaques in cell culture is associated with suppression of alpha/beta
interferon. J Virol 73, 9891-9898 (1999).
Cho, J.G. et al. Evaluation of the effects of animal age, concurrent bacterial infection,
and pathogenicity of porcine reproductive and respiratory syndrome virus on virus
concentration in pigs. Am J Vet Res 67, 489-493 (2006).
Chung, H.K., Lee, J.H., Kim, S.H. & Chae, C. Expression of interferon-alpha and Mx1
protein in pigs acutely infected with porcine reproductive and respiratory syndrome
virus (PRRSV). J Comp Pathol 130, 299-305 (2004).
Clemens, M.J. & Elia, A. The double-stranded RNA-dependent protein kinase PKR:
structure and function. J Interferon Cytokine Res 17, 503-524 (1997).
Conzelmann, K.K., Visser, N., Van Woensel, P. & Thiel, H.J. Molecular
characterization of porcine reproductive and respiratory syndrome virus, a member of
the arterivirus group. Virology 193, 329-339 (1993).
Crance, J.M., Scaramozzino, N., Jouan, A. & Garin, D. Interferon, ribavirin,
6-azauridine and glycyrrhizin: antiviral compounds active against pathogenic
flaviviruses. Antiviral Res 58, 73-79 (2003).
DeLano, W.L., Ultsch, M.H., de Vos, A.M. & Wells, J.A. Convergent solutions to
binding at a protein-protein interface. Science 287, 1279-1283 (2000).
Diebold, S.S., Kaisho, T., Hemmi, H., Akira, S. & Reis e Sousa, C. Innate antiviral
responses by means of TLR7-mediated recognition of single-stranded RNA. Science
303, 1529-1531 (2004).
Doucey, M.A., Hess, D., Cacan, R. & Hofsteenge, J. Protein C-mannosylation is
enzyme-catalysed and uses dolichyl-phosphate-mannose as a precursor. Mol Biol Cell
9, 291-300 (1998).
Edwards, C.P. & Aruffo, A. Current applications of COS cell based transient
expression systems. Curr Opin Biotechnol 4, 558-563 (1993).
Fornwald, J.A. et al. Soluble forms of the human T cell receptor CD4 are efficiently
expressed by Streptomyces lividans. Biotechnology (N Y) 11, 1031-1036 (1993).
Franco, R., Daniela, G., Fabrizio, M., Ilaria, G. & Detlev, H. Influence of osmolarity and pH increase to achieve a reduction of monoclonal antibodies aggregates in a
production process. Cytotechnology 29, 11-25 (1999).
Freedman, R.B. Protein disulfide isomerase: multiple roles in the modification of
nascent secretory proteins. Cell 57, 1069-1072 (1989).
Garcia-Sastre, A. et al. The role of interferon in influenza virus tissue tropism. J Virol
72, 8550-8558 (1998).
George, C.X. & Samuel, C.E. Human RNA-specific adenosine deaminase ADAR1
transcripts possess alternative exon 1 structures that initiate from different promoters,
one constitutively active and the other interferon inducible. Proc Natl Acad Sci U S A
96, 4621-4626 (1999).
Ghetie, V. & Ward, E.S. Multiple roles for the major histocompatibility complex class
I- related receptor FcRn. Annu Rev Immunol 18, 739-766 (2000).
Gillies, S.D., Young, D., Lo, K.M. & Roberts, S. Biological activity and in vivo
clearance of antitumor antibody/cytokine fusion proteins. Bioconjug Chem 4, 230-235
(1993).
Glue, P. et al. Pegylated interferon-alpha2b: pharmacokinetics, pharmacodynamics,
safety, and preliminary efficacy data. Hepatitis C Intervention Therapy Group. Clin
Pharmacol Ther 68, 556-567 (2000).
Guan, K.L. & Dixon, J.E. Eukaryotic proteins expressed in Escherichia coli: an
improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem 192, 262-267 (1991).
Haller, O., Frese, M. & Kochs, G. Mx proteins: mediators of innate resistance to RNA
viruses. Rev Sci Tech 17, 220-230 (1998).
Harris, M. Interferon: clinical application of molecular biology. Science 170,
1068-1070 (1970).
Heil, F. et al. Species-specific recognition of single-stranded RNA via toll-like
receptor 7 and 8. Science 303, 1526-1529 (2004).
Helenius, A. & Aebi, M. Roles of N-linked glycans in the endoplasmic reticulum.
Annu Rev Biochem 73, 1019-1049 (2004).
Hitzeman, R.A. et al. Use of heterologous and homologous signal sequences for
secretion of heterologous proteins from yeast. Methods Enzymol 185, 421-440 (1990).
Hofsteenge, J., Blommers, M., Hess, D., Furmanek, A. & Miroshnichenko, O. The
four terminal components of the complement system are C-mannosylated on multiple
tryptophan residues. J Biol Chem 274, 32786-32794 (1999).
Huang, T., Pavlovic, J., Staeheli, P. & Krystal, M. Overexpression of the influenza
virus polymerase can titrate out inhibition by the murine Mx1 protein. J Virol 66,
4154-4160 (1992).
Hwang, S.Y. et al. A null mutation in the gene encoding a type I interferon receptor
component eliminates antiproliferative and antiviral responses to interferons alpha andbeta and alters macrophage responses. Proc Natl Acad Sci U S A 92, 11284-11288(1995).
Isaacs, A. & Lindenmann, J. Virus interference. I. The interferon. Proc R Soc Lond B
Biol Sci 147, 258-267 (1957).
Jayapal, K.P. et al. Recombinant protein therapeutics from CHO cells – 20 years and
counting. Chem. Eng. Prog. 103, 40–47 (2007).
Jazayeri, J.A. & Carroll, G.J. Fc-based cytokines : prospects for engineering superior
therapeutics. BioDrugs 22, 11-26 (2008).
Jenkins, N. & Curling, E.M. Glycosylation of recombinant proteins: problems and
prospects. Enzyme Microb Technol 16, 354-364 (1994).
Jenkins, N., Parekh, R.B. & James, D.C. Getting the glycosylation right: implications
for the biotechnology industry. Nat Biotechnol 14, 975-981 (1996).
Jiang, Z., Mak, T.W., Sen, G. & Li, X. Toll-like receptor 3-mediated activation of
NF-kappaB and IRF3 diverges at Toll-IL-1 receptor domain-containing adapter
inducing IFN-beta. Proc Natl Acad Sci U S A 101, 3533-3538 (2004).
Johansson, E., Domeika, K., Berg, M., Alm, G.V. & Fossum, C. Characterisation of
porcine monocyte-derived dendritic cells according to their cytokine profile. Vet
Immunol Immunopathol 91, 183-197 (2003).
Jones, T.D. et al. The development of a modified human IFN-alpha2b linked to the Fc
portion of human IgG1 as a novel potential therapeutic for the treatment of hepatitis C
virus infection. J Interferon Cytokine Res 24, 560-572 (2004).
Kaufman, R.J. Selection and coamplification of heterologous genes in mammalian
cells. Methods Enzymol 185, 537-566 (1990).
Kerr, I.M. The 2-5A system: a personal view. J Interferon Res 7, 505-510 (1987).
Kim, J.K., Tsen, M.F., Ghetie, V. & Ward, E.S. Localization of the site of the murine
IgG1 molecule that is involved in binding to the murine intestinal Fc receptor. Eur J
Immunol 24, 2429-2434 (1994).
Kitts, P.A. & Possee, R.D. A method for producing recombinant baculovirus
expression vectors at high frequency. Biotechniques 14, 810-817 (1993).
Kornfeld, R. & Kornfeld, S. Assembly of asparagine-linked oligosaccharides. Annu
Rev Biochem 54, 631-664 (1985).
Kozak, M. At least six nucleotides preceding the AUG initiator codon enhance
translation in mammalian cells. J Mol Biol 196, 947-950 (1987).
Kukuruzinska, M.A., Bergh, M.L. & Jackson, B.J. Protein glycosylation in yeast. Annu
Rev Biochem 56, 915-944 (1987).
Li, J. et al. Preparation of CHO cell-derived rhIFN-omega-Fc with improved
pharmacokinetics. Antiviral Res 89, 199-203 (2011).
Lo, K.M. et al. High level expression and secretion of Fc-X fusion proteins in
mammalian cells. Protein Eng 11, 495-500 (1998).
Luo, R. et al. Antiviral activity of type I and type III interferons against porcine
reproductive and respiratory syndrome virus (PRRSV). Antiviral Res 91, 99-101
(2011).
Maina, C.V. et al. An Escherichia coli vector to express and purify foreign proteins by
fusion to and separation from maltose-binding protein. Gene 74, 365-373 (1988).
Makrides, S.C. Components of vectors for gene transfer and expression in mammalian
cells. Protein Expr Purif 17, 183-202 (1999).
Marston, F.A. & Hartley, D.L. Solubilization of protein aggregates. Methods Enzymol
182, 264-276 (1990).
Matsuura, Y., Possee, R.D., Overton, H.A. & Bishop, D.H. Baculovirus expression
vectors: the requirements for high level expression of proteins, including glycoproteins.
J Gen Virol 68 ( Pt 5), 1233-1250 (1987).
Meulenberg, J.J. et al. Lelystad virus, the causative agent of porcine epidemic abortion
and respiratory syndrome (PEARS), is related to LDV and EAV. Virology 192, 62-72
(1993).
Muller, U. et al. Functional role of type I and type II interferons in antiviral defense.
Science 264, 1918-1921 (1994).
Ober, R.J., Martinez, C., Vaccaro, C., Zhou, J. & Ward, E.S. Visualizing the site and
dynamics of IgG salvage by the MHC class I-related receptor, FcRn. J Immunol 172,
2021-2029 (2004).
Okuse, C., Rinaudo, J.A., Farrar, K., Wells, F. & Korba, B.E. Enhancement of
antiviral activity against hepatitis C virus in vitro by interferon combination therapy.
Antiviral Res 65, 23-34 (2005).
Overend, C. et al. Recombinant swine beta interferon protects swine alveolar
macrophages and MARC-145 cells from infection with Porcine reproductive and
respiratory syndrome virus. J Gen Virol 88, 925-931 (2007).
Pavlovic, J., Haller, O. & Staeheli, P. Human and mouse Mx proteins inhibit different
steps of the influenza virus multiplication cycle. J Virol 66, 2564-2569 (1992).
Pavlovic, J., Schroder, A., Blank, A., Pitossi, F. & Staeheli, P. Mx proteins: GTPases
involved in the interferon-induced antiviral state. Ciba Found Symp 176, 233-243;
discussion 243-237 (1993).
Pestka, S., Langer, J.A., Zoon, K.C. & Samuel, C.E. Interferons and their actions.
Annu Rev Biochem 56, 727-777 (1987).
Pestka, S., Krause, C.D. & Walter, M.R. Interferons, interferon-like cytokines, and
their receptors. Immunol Rev 202, 8-32 (2004).
Peters, B.P., Krzesicki, R.F., Perini, F. & Ruddon, R.W. O-glycosylation of the
alpha-subunit does not limit the assembly of chorionic gonadotropin alpha beta dimer in human malignant and nonmalignant trophoblast cells. Endocrinology 124,
1602-1612 (1989).
Piechaczek, C., Fetzer, C., Baiker, A., Bode, J. & Lipps, H.J. A vector based on the
SV40 origin of replication and chromosomal S/MARs replicates episomally in CHO
cells. Nucleic Acids Res 27, 426-428 (1999).
Pitha, P.M., Wivel, N.A., Fernie, B.F. & Harper, H.P. Effect of interferon on murine
leukaemia virus infection. IV. Formation of non-infectious virus in chronically
infected cells. J Gen Virol 42, 467-480 (1979).
Pitossi, F. et al. A functional GTP-binding motif is necessary for antiviral activity of
Mx proteins. J Virol 67, 6726-6732 (1993).
Player, M.R. & Torrence, P.F. The 2-5A system: modulation of viral and cellular
processes through acceleration of RNA degradation. Pharmacol Ther 78, 55-113
(1998).
Proud, C.G. PKR: a new name and new roles. Trends Biochem Sci 20, 241-246 (1995).
Rademacher, T.W., Parekh, R.B. & Dwek, R.A. Glycobiology. Annu Rev Biochem 57,
785-838 (1988).
Raghavan, M., Chen, M.Y., Gastinel, L.N. & Bjorkman, P.J. Investigation of the
interaction between the class I MHC-related Fc receptor and its immunoglobulin G
ligand. Immunity 1, 303-315 (1994).
Raghavan, M., Wang, Y. & Bjorkman, P.J. Effects of receptor dimerization on the
interaction between the class I major histocompatibility complex-related Fc receptor
and IgG. Proc Natl Acad Sci U S A 92, 11200-11204 (1995).
Raghavan, M. & Bjorkman, P.J. Fc receptors and their interactions with
immunoglobulins. Annu Rev Cell Dev Biol 12, 181-220 (1996).
Rai, M., Padh, H., Expression systems for the production of heterologous proteins.
Curr Sci 80:1121–1128 (2001)
Renault T., Torchy C., De Kinkelin P. Spectrophotometric method for titration of trout
interferon, and its application to rainbow trout fry experimentally infected with viral
hemorrhagic epticemia virus. Dis. Aquat. Organ. 10:23–9 (1991).
Robinson, M. et al. Codon usage can affect efficiency of translation of genes in
Escherichia coli. Nucleic Acids Res 12, 6663-6671 (1984).
Roopenian, D.C. & Akilesh, S. FcRn: the neonatal Fc receptor comes of age. Nat Rev
Immunol 7, 715-725 (2007).
Rossow, K.D. Porcine reproductive and respiratory syndrome. Vet Pathol 35, 1-20
(1998).
Rudd, P.M., Elliott, T., Cresswell, P., Wilson, I.A. & Dwek, R.A. Glycosylation and
the immune system. Science 291, 2370-2376 (2001).
Samuel, C.E. Antiviral actions of interferon. Interferon-regulated cellular proteins and their surprisingly selective antiviral activities. Virology 183, 1-11 (1991).
Sarkar, S.N. & Sen, G.C. Production, purification, and characterization of recombinant
2', 5'-oligoadenylate synthetases. Methods 15, 233-242 (1998).
Schriebl, K. et al. Biochemical characterization of rhEpo-Fc fusion protein expressed
in CHO cells. Protein Expr Purif 49, 265-275 (2006).
Sharma, S. et al. Triggering the interferon antiviral response through an IKK-related
pathway. Science 300, 1148-1151 (2003).
Simister, N.E. & Mostov, K.E. An Fc receptor structurally related to MHC class I
antigens. Nature 337, 184-187 (1989).
Smith, G.E., Summers, M.D. & Fraser, M.J. Production of human beta interferon in
insect cells infected with a baculovirus expression vector. Mol Cell Biol 3, 2156-2165(1983).
Song, X.Y., Torphy, T.J., Griswold, D.E. & Shealy, D. Coming of age: anti-cytokine
therapies. Mol Interv 2, 36-46 (2002).
Spiegelberg, H.L. & Fishkin, B.G. The catabolism of human G immunoglobulins of
different heavy chain subclasses. 3. The catabolism of heavy chain disease proteins
and of Fc fragments of myeloma proteins. Clin Exp Immunol 10, 599-607 (1972).
Staeheli, P., Dreiding, P., Haller, O. & Lindenmann, J. Polyclonal and monoclonal
antibodies to the interferon-inducible protein Mx of influenza virus-resistant mice. J Biol Chem 260, 1821-1825 (1985).
Staeheli, P. & Pavlovic, J. Inhibition of vesicular stomatitis virus mRNA synthesis by
human MxA protein. J Virol 65, 4498-4501 (1991).
Staeheli, P., Pitossi, F. & Pavlovic, J. Mx proteins: GTPases with antiviral activity.
Trends Cell Biol 3, 268-272 (1993).
Stark, G.R., Kerr, I.M., Williams, B.R., Silverman, R.H. & Schreiber, R.D. How cells
respond to interferons. Annu Rev Biochem 67, 227-264 (1998).
Suradhat, S., Intrakamhaeng, M. & Damrongwatanapokin, S. The correlation of
virus-specific interferon-gamma production and protection against classical swine
fever virus infection. Vet Immunol Immunopathol 83, 177-189 (2001).
Takeda, K. & Akira, S. TLR signaling pathways. Semin Immunol 16, 3-9 (2004).
Tharmalingam, T., Sunley, K. & Butler, M. High yields of monomeric recombinant
beta-interferon from macroporous microcarrier cultures under hypothermic conditions.
Biotechnol Prog 24, 832-838 (2008).
Trill, J.J., Shatzman, A.R. & Ganguly, S. Production of monoclonal antibodies in COS
and CHO cells. Curr Opin Biotechnol 6, 553-560 (1995).
Van der Linden, I.F., Voermans, J.J., van der Linde-Bril, E.M., Bianchi, A.T. &
Steverink, P.J. Virological kinetics and immunological responses to a porcine
reproductive and respiratory syndrome virus infection of pigs at different ages.
Vaccine 21, 1952-1957 (2003).
Vaughn, D.E. & Bjorkman, P.J. Structural basis of pH-dependent antibody binding by
the neonatal Fc receptor. Structure 6, 63-73 (1998).
Vieira, P. & Rajewsky, K. The half-lives of serum immunoglobulins in adult mice. Eur
J Immunol 18, 313-316 (1988).
Wang, Y.B. et al. Secretory expression of porcine interferon-gamma in baculovirus
using HBM signal peptide and its inhibition activity on the replication of porcine
reproductive and respiratory syndrome virus. Vet Immunol Immunopathol 132,
314-317 (2009).
Wesley, R.D., Lager, K.M. & Kehrli, M.E., Jr. Infection with Porcine reproductive and
respiratory syndrome virus stimulates an early gamma interferon response in the serum
of pigs. Can J Vet Res 70, 176-182 (2006).
West, A.P., Koblansky, A.A. & Ghosh, S. Recognition and signaling by toll-like
receptors. Annu Rev Cell Dev Biol 22, 409-437 (2006)
Wiebe, M. E., et al., "A multifaceted approach to assure that recombinant tPA is free of
adventitious virus," Advances in animal cell biology and technology,
Butterworth-Heinemann, London, pp. 68-71 (1989).
Williams, R.J. Calcium binding proteins in normal and transformed cells, Perugia,
May 1996. Cell Calcium 20, 87-93 (1996).
Yoon, S.K., Kim, S.H. & Lee, G.M. Effect of low culture temperature on specific
productivity and transcription level of anti-4-1BB antibody in recombinant Chinese
hamster ovary cells. Biotechnol Prog 19, 1383-1386 (2003).
Zhang, X., Shin, J., Molitor, T.W., Schook, L.B. & Rutherford, M.S. Molecular
responses of macrophages to porcine reproductive and respiratory syndrome virus
infection. Virology 262, 152-162 (1999).
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1. 50.葉石濤《臺灣文學史綱》,高雄:文學界雜誌社,1987年2月。
2. 顏妙桂 (2002),九年一貫課程綱要休閒教育內涵之探討-從能力指標分析,公民訓育學報,12 輯,頁33-58。
3. 12.司徒衛:〈五十年代自由中國的新文學〉,《文訊》9期,臺北:文訊雜誌出版社,1984年。
4. 黃致傑,陳鎰明(2003),休閒教育在國小的實施之探討,雲科大體育,6 期,頁104-110。
5. 黃立賢(1999),中小學教師實施休閒教育實務篇──觀念與政策之探討。台北:學生輔導雙月刊,60 期。pp80∼89。
6. 張景然 (1991),休閒生活與青少年行為輔導,成人教育,2 期,頁48-49。
7. 1.上官予:〈五十年代的新詩〉,《文訊》9期,1984年3月。
8. 6.江寶釵:〈與文學傳媒結緣-談臺灣新文學期刊的研究〉,《文訊雜誌》,第213期,2003年。
9. 7.江寶釵:〈重省五○年代臺灣文學史的詮釋問題──一個奠基於「場域」的思考〉,《東華漢學》3期,東華大學中國語文學系,2005年5月。
10. 8.何義麟:〈戰後初期臺灣出版事業之傳承與移植(1945~1950)-雜誌目錄初編後之考察〉,《臺灣史料研究》,第十期,1997年12月。
11. 9.李牧:〈新文學運動歷程中的關鍵時代─試探五0年代自由中國文學創作的思路及其所產生的影響〉,《文訊》,第9期,1984年3月。
12. 13.封德屏〈精神與風格的展現—文學雜誌的專題設計〉,《文訊雜誌》,第27期,1986年12月。
13. 14.栗國成:〈1957年臺北「劉自然事件」及 1965 年〈美軍在華地位協定〉之簽訂〉:東吳政治學報,2006年。
14. 15.張素貞:〈五十年代小說管窺〉,《文訊》,第9期,1984年3月。
15. 16.張淑雅:〈一九五○年代美國對臺決策模式分析〉,《中央研究院近代史研究所集刊》,第40期, 2003年6月。