跳到主要內容

臺灣博碩士論文加值系統

(3.215.79.68) 您好!臺灣時間:2022/07/04 04:22
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:白佳真
研究生(外文):Chia -Chen Pai
論文名稱:樟芝免疫調節蛋白與超氧岐化酶之研究
論文名稱(外文):Studies on the Fungal Immunomodulatory Protein and Superoxide Dismutase from Antrodia camphorata
指導教授:林棋財
指導教授(外文):Chi-Tsai Lin
學位類別:碩士
校院名稱:國立海洋大學
系所名稱:生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2003
畢業學年度:91
語文別:中文
論文頁數:96
中文關鍵詞:樟芝免疫調節蛋白超氧歧化酶鍊聚合酶反應
外文關鍵詞:Antrodia camphorataFungal immunomodulatory proteinSuperoxide dismutasePolymerase chain reaction
相關次數:
  • 被引用被引用:10
  • 點閱點閱:1207
  • 評分評分:
  • 下載下載:195
  • 收藏至我的研究室書目清單書目收藏:3
牛樟芝 ( Antrodia camphorata )為一台灣特有菌種,生長於牛樟樹 ( Cinnamomun kanehirai )中空樹洞中,會造成牛樟樹產生褐腐病(brown rot disease)。據民俗療法的記載,牛樟芝具有保肝解毒的能力,其保肝的機轉目前已知與免疫調節蛋白與抗氧化酵素的功能有極大的關係。
藉由聚合酶連鎖反應( PCR ),進行樟芝Fips與SODs的cDNA與基因選殖,並進行蛋白質表現。依據松杉靈芝、草菇、金針菇的Fips胺基酸序列,在其保守區域設計7組primers,經由多次不同條件的PCR及次選殖之後,目前尚未發現與Fips類似的序列。
在SOD選殖方面,在樟芝的子實體中選殖出3條SOD基因的部分片段,分別命名為AcaMn SOD I、AcaMn SOD II 及AcaFe SOD 。AcaMn SOD I 為genomic DNA,長度為420 bp,可轉譯出140 個胺基酸;AcaMn SOD II 為樟芝cDNA序列,長度為489 bp,可轉譯為105個胺基酸;AcaFe 為樟芝cDNA序列,長度為423 bp,可轉譯出129個胺基酸。
此外,自中研院蕭介夫老師實驗室所建構的樟芝菌絲體EST中取得一條全長Mn SOD的cDNA序列,其全長為954 b.p.,coding region 為693 bp,可轉譯出230 個胺基酸,命名為AcaMn SOD III 。以樟芝子實體的cDNA為模板,依據AcaMn SOD III表現區序列合成2段primers ( AcaMn-a, AcaMn-b )進行PCR,可得到與AcaMn SOD III中相同的Mn SOD 表現區。經由胺基酸比對,得知其與靈芝屬菌類及白枯真菌皆有很高的相似性,並且具有與文獻中所述Mn 離子結合有關的4個胺基酸( His59, His104, Asp189 , His193 ) 與活性區8個胺基酸中提供鍵結形成dimer型式的2個胺基酸 ( Glu192, Tyr197 ) 。將之接在pET-20b(+)中,以BL21(DE3 )pLysS為表現宿主,在C-端未接His-tag表現的plasmid中,可表現具有活性的SOD。另以樟芝子實體或菌絲體genomic DNA為模板,分別與AcaMn-a及AcaMn-b primer進行PCR,兩者均可得到具有4個 intron的SOD基因,且分析其基因序列完全相同。這些結果有助於未來進一步探討Fips與SODs。
Antrodia camphorata one kind of the medicinal mushrooms, can naturally parasite in the punk of Cinnamomun kanehirai Hay. In ethnologic medicine, A. camphorata has the activation of detoxification, anticancer and curing liver complaint. Evidences have showed that Fip and SOD are known on the physiologically active component in mushrooms. This motivates we to clone DNA of Fip and SOD from A. camphorata.
In accordance with the conserve amino acid of Fip or SOD from other species, we try to clone DNA of Fip and SOD from A. camphorata,3 clones of SOD (AcaMn SOD I, AcaMn SOD II, AcaFe SOD) were screened. AcaMn SOD I is a genomic DNA contains 420 bp, and its open reading frame ecoding 140 amino acid residues; AcaMn SOD II is a cDNA contains 489 bp, and its open reading frame ecoding 105 amino acid residues. AcaFe SOD is a cDNA contains 423 bp, and open reading frame ecoding 129 amino acid residues.
Meanwhile, another full-length Mn SOD sequence was obtained from A. camphorata mycelia (kindly given from Dr. Shaw’s Lab) named AcaMn SOD III containing 954 bp, and its open reading frame ecoding 230 amino acid residues. According to AcaMn SOD III sequence, 2 primers were synthesized, when using A. camphorata fruit bodies cDNA as a template, an 0.6 Kb coding region was amplified by PCR. Computer analysis of the residues required for coordinating the single trivalent manganese iron and 11 residues putatively involve in the active center were well conserve among all reported Mn SOD. Whereas, using A. camphorata genomic DNA as a template, an 0.8 Kb was amplified .Sequence analysis revealed that 4 introns were intervened in the AcaMn SOD III.
To furthermore characterize the A. camphorata Mn SOD, the coding region was subcloned into BL21(DE3)pLysS. The active SOD was expressed of the recombinat protein which no His-tag at C-terminal.
目錄
中文摘要……………………………………………………………. II
英文摘要…………………………………………………………….IV
縮寫表………………………………………………………………..V
壹、 前言………………………………………………………… … 1
貳、 實驗材料與方法……………………………………………….33
參、 結果與討論…………………………………………………….58
參考文獻…………………………………………………………….83
附錄………………………………………………………………….88
陳勁初、林文鑫、陳勁農、許勝傑、黃仕政、陳炎鍊..台灣特有真菌-樟芝菌絲體之開發。Fung. Sci. 2001, 16(1,2):7-22。
王伯徹、陳啟楨、華傑. 食藥用菇類的培養與應用. 食品工業發展研究所: 第M-87-019號. 1998,新竹,台灣. 187頁.
王伯徹、黃仁彰. 靈芝與樟芝之研發與是場面面觀. 食品工業2002, 349(5): 3-17.
.李學勇. 基礎植物學. 1990,台北市 國立編譯館
易希珍. 普通植物學. 1976,台北市 環球出版社
山羽儀兵 釋譯者.三版. 植物通論. 1976,台北市. 正中出版社
張東柱、謝煥儒、張瑞璋、傅春旭. 1999,台灣常見樹木病害
Ames, B.; Shigenaga, K. and Hagen, T. M. Oxidants, antioxidants and the degenerative diseases of aging. Proc. Natl. Acad. Sci. USA. 1993, 90: 7915-7922.
Bannister, J. V.; Bannister, W. H. and Rotilio, G. Aspect of the structure, funtion and applications of superoxide dismutase. Crit. Rev. Biochem. 1987, 22: 111-112.
Bridges, S. M. Distribution of iron containing superoxide dismutase in vesular plants. Plant Physiol. 1981, 68: 275-278.
Bourgeois, C.; Corvaisier, C.,B.; Kohli, E. and Pothier, P.Use of synthetic peptides to locate neutralizing anti-genic demains on the fusion protein of respiratory syncytial virus. J Gen. Virol. 1991, 72:1051-1058.
Cadenas, E. Mechanisms of oxygen activation and reactive oxygen species detoxification. Oxidative Stress and Antioxidant Defenses in Biol. 1995, 1-61.
Cavalieri, E. L. and Rogan, E. G. Central role of radical cations in metabolic activation of polycyclic aromatic hydrocarbons. Xenobiotica. 1995, 25: 677-688.
Celada, A.; Gray, P. W.; Rinderknecht, F. and Scheber, R. D.Evidence for a gamma interferon receptor that regulates macrophage tumoricidal activity. J. Exp. Med. 1984, 160:55.
Chang, T. T. and W. N. Chou.. Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycol. Res. 1995, 99: 756-758.
http://www.hkjh.tc.edu.tw/~bio/newpage31.htm
Chen, C. H.; Yang, S. W. .New steroid acid from Antrodia xinnamomea a fungal parasite of Cinnanomun micranthum.J. Nat. Prod. 1995, 58:1655-1661.
Cheng, K.; Cahill, H. and Kasai, S. 8-Hydroxyguanine, an abundant from of oxidative DNA damage, causes GT and AC substitution. J. Biol. Chem. 1992, 267: 166-172.
Church, D.; Pryor, F. A. and Pryor, W. A. Free radical chemistry of cigarette smoke and its toxicological implications. Environ. Health Perspect. 1985, 64: 111-126.
Curnutte, J. T. and Babior, B. M. Chronic granulomatous disease. Adv. Hum.Genet. 1987, 16: 229-297.
Elizabeth, C. and Daniel, E. NO news is good news. Science. 1992, 258(18):1862-1865.
Frankel, E. N. Recent advances in lipid oxidation. J. Sci. Food Agric. 1991, 54: 495-511.
Fridovich, I. Superoxide dismutases. Adv. Enzymol. 1986, 58: 61-97.
Green, M. J. and Hill, H. A. Chemistry of dioxygen. Methods Enzymol. 1984, 105: 3-22.
Hagen, U. Biochemical aspects of radiation biology. Experientia. 1989, 45(1): 7-12.
Halliwell, B.; Murcia, M. A.; Chirico, S. and Aruoma, O. I. Free radicals and antioxidants in food and in vivo: what they do and how they work. Crit. Rev. Food Sci. Nutr. 1995, 35: 7-20.
Halliwell, B. The Lancet. 1994, 344: 721-724.
Hallwell B. The role of oxygen radicals in human disease, with particular reference to the vascular system. Haemostasis. 1993, 23: 118-126.
Hazlewood, C. and Davies, M. J. Damage to DNA and RNA by tumour promoter-derived alkoxyl radicals: an EPR spin trapping study. Biochem. Soc. Trans 1995, 23: 259-260.
Hseu, Y. C.; Chang, W. C.; Hseu, Y. T.; Lee, C. Y.;Yech, Y. J.; Chen, P. C. ; Chen, J. Y.; Yang, H. L. Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in nomal human erythrocytes.Life Sci. 2002, 71:469-482.
Hsiao, G. ; Shen, M. Y. ; Lin, K. H.; Lan, M. H. ; Wu, L. Y.; Chou, D. S.; Su, C. H. and Sheu, J. R. Antioxidative and Hepatoprotective effect of Antrodia camphorata extract.J. Agric. Food Chem. 2003, 51:3302-3308.
Huber, R. and Warner. Superoxide dismutase, aging and degenerative disease. Free radical Biol. Med. 1994, 17: 249-258.
Hsu, H. C.; Hsu, C. I.; Lin, R.H. ; Kao, C. L. and Lin, J. Y. Fip-vvo, a new fungal immunomodulatory protein isolated from Volvariella volvacea. Biochem. J. 1997,.323:557-565
James, P. and Kehrer, D. Free radicals as mediators of tissue injury and disease. Critical Reviews in Toxicology. 1993, 23(1): 21-48.
Jonathan, S.; David, J. and Singel, J. L. Biochemistry of nitric oxide and its redox-activated forms. Science. 1992, 258(18): 1898-1902.
Kawagishi, H. ; Mitsunaga, S. I.; Yamawaki, M. ; Ido, M.; Shimada, A. ; Tetsuya, Murata, T.; Usui, T.; Kimura, A. and Chiba. A lectin from mycelia of fungus Ganoderma lucidum.Phytochemistry. 1997, 44(1):7-10.
Kino, K. ; Yamashita, A .; Yamaoka , K.; Watanabe, J.; Tanaka, S.; Ko, K.; Shimizu, K. and Tsunoo, H. Isolation and characterization of a new immunomodulatory protein, Ling Zhi-8 ( LZ-8 ),from Ganoderma lucidium. J. Biol. Chem. 1989, 1:472-478.
Ko, J. L; Hsu, C. I.; Lin, R. H.; Kao, C. L. and Lin, J. Y. A new fungal immunomodulatory protein, FIP-fve isolated the edible mushroom Flammulina velitipes and its complete amino acid sequence.Eur. J. Biochem 1995, 228:244-249.
Lee, I. H. ; Huang, R. L.; Chen, C. T.; Chen, H. C.; Hsu, W. C.; Lu, M. K. .Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects.FEMS Microbiology letters 2002, 209:63-67.
Lim, J. H.; Yu, Y. G.; Han, Y. S.; Cho, S. J.; Ahn, B. Y.; Kim, S. H. and Cho, Y. The crystal structure of an Fe-superoxide dismutase from the hyperthermophile Aquifex pyrophilius at 1.9Å resolution: structure basis for thermstability. J. Mol. Biol. 1997, 270: 259-274.
Lin, C. T.; Kuo, T. J.; Shaw, J. F. and Kao, M. C. Characterization of the dimer-monomer equilibrium of the papaya copper/zinc superoxide dismutase and it’s equilibrium shift by a single amino acid mutation. J. Arg. Food.Chem. 1999, 47: 2944-2949.
Lin, M. T.;Kuo, T. J. and Lin, C. T. Molecular cloning of a cDNA encoding copper/zinc superoxide dismutase from papaya fruit and overexpression in E.coli. J.Agric.Food Chem. 1998, 46: 344-348.
Lin, W. H.; Hung, C. H.; Hsu, C. I. and Lin, J. Y. . Dimerization of the N-terminal Amphipathic a-Helix Domain of the Fungal Immunomodulatory Protein from Ganoderma tsugae(Fip-gts) Defined by a Yeast Two-hybrid System and
Site-directed Mutagenesis. J. Biol. Chem 1997, 22:20044-20048.
Maly, F. E. The β-lymphocyte: a newly recognized source of reactive oxygen species with immunoregulatory potential. Free Rad. Res. Commun. 1990, 8: 143-148.
Mann, T. and Keilin, D. Haemocuprein and hepatocuprein, copper-proein compounds of blood and liver in mammals. Proc. R. Soc. London Ser B. 1939, 128: 303-304.
McCord, J. M. and Fridovich, I. Superoxide dismutase. An enzymatic function for erythrocuprein (hemocuprein). J. Biol. Chem. 1969, 244: 6049-6052.
Meier, B.; Radeke, H.; Selle, S.; Raspe, H. H.; Sies, H.; Resch, K. and Habermehl, G. G. Human fibroblasts release reactive oxygen species in response to treatment with synovial fluids from patients suffering from arthritis. Free Radical Res. Commun. 1990, 8: 149-160.
Narumi, S.; Finke, J. H. and Hamilton, T. A. IFN-γ and IL-2synergize to induce selective monokine expression in murine peritoneal macrophages. J. Biol. Chem. 1990, 265:7036.
Nathan, C. F. and Hibbs. J. B., Jr. Role of nitric oxide synthesis in macrophage antimicrobial activity. Curr.Opin. Immunol. 1991., 3:65.
Pace, L.; Russel, S. W.; Torres, B. A.,; Johnson, H. M. and Gray, P. W..Recombinant mouseγ interferon induces the primimg step in macrophage activation for tumor cell killing.J. Immunol. 1983, 130:2011.
Pan, S. M. and Y. Y. Yan. The isozymes of superoxide dismutase in rice. Bot. Bull. Acad. Sin. 1991, 32: 253-258.
Pryor, W. A., Lightsey, J. W. and Church, D. F. Reaction of nitrogen dioxide with alkenes and polyunsaturated fatty acids: addition and hydrogen abstraction mechanisms. J. Am. Chem. Soc. 1982, 104: 6685-6671.
Rafael, R.; Marianela, R.; Laura, C. and Rossana, T. Inhibition of mitochondrial electron transport by peroxynitrite. Arch. Biochem. Biophy. 1994, 308: 89-95.
Reid, T. M. and Loeb, L. A. Effect of DNA repair enzymes on mutagenesis by oxygen free radicals. Mutat. Res 1993, 289: 181-186.
Sandy, M. S.; Moldeus, P.; Ross, D. and Smith, M. T.. Role of redox cycling and lipid peroxidation in bipyridyl herbicide cytotxicity. Studies with a compromised isolated hepatocyte model system. Biochem. Pharmacol. 1986, 35: 3095-3101.
Schinina, M.E., Carlini, P., Polticelli, F., Zappacosta, F., Bossa, F. and Calabrese L. Amino acid sequence of chicken Cu,Zn-containing superoxide dismutase and identification of glutathionyl adducts at exposed cysteine residues. Euro. J. Biochem. 1996, 237(2): 433-439.
Simic, M. G. Mechanisms of inhibition of free-radical processes in mutagenesis and carcinogenesis. Mutat. Res. 1988,. 202: 377-386.
Stadtman, E. R. and Oliver, C. N. Metal-catalyzed oxidation of proteins. Physiological consequences. J. Biol. Chem. 1991, 266: 2005-2008.
Stadtman, E. R. Protein oxidation and aging. Science. 1992,257: 1220-1224
Steinman, H. M.; Naik, U. R.; Aberenthy, J. L. and Hill, R. A. Bovine erythrocyte superoxide dismutase. Complete amino acid sequence. J. Biol. Chem. 1974, 249: 7326-7338.
Song, T. Y. and Yen, G. C. Antioxidant properties of Antrodia camphorata in submerged culture. J. Agirc. Food Chem. 2002, 50:3322-3327.
Song, T. Y. and Yen, G. C. Protective effect of fermented filtrate from Antrodia camphorata in Submerged culture against CCl4-induced hepatic toxicity in rat. J. Agric. Food Chem. 2003, 51:1571-1577.
Tainer, J. A.; Getzoff, E. D.; Beem, K. M. and Richardson, D. Structure and mechanism of copper, zinc superoxide dismutase. Nature. 1983, 306: 284-286.
Tsai, Z. T.;Liaw, S. L..The use and the effect of Ganoderma .San Yun Press: Taichung , Taiwan 1985 , 116.
Tanaka, S.; Ko, K.; Kino, K. ; Tsuchiya, K. ; Yamashita, A. ; Murasugi, A. ; Sakuma, S. and Tsunoo, H. Complete amino acid sequence of immunomodulatory protein,Ling Zhi(LZ-8). J. Biol. Chem. 1989, 264:16372-16377.
Wayne, F.; Beyer, J. and Fridovich, I. In vivo competition between iron and manganese for occupancy of the active site region of the manganese-superoxide dismutase of Escherichia coli. J. Biol. Chem. 1991, 266 (1): 303-315.
William, C. S.; Katherine, A. P.; Roland, K. S. and Martha, L. L. Manganese and iron superoxide dismutase are structural homology. J. Biol. Chem. 1984, 259 (17): 10695-10702.
Van der Hem, L. G.; Van der Vilet, J. A., Bokem, C. F. M., Kino, K., Hoitsma, A. J. and Tax, W. J. M.. LZ-8: Studies of a new immnomodulatory agent. Transplatation 1995, 60:438-443.
Yan, T.; Larry, W. O. and Weixiong, Z. Manganese-containing superoxide dismutase overexpression causes phenotypic reversion in SV40-transfomed human lung fibroblasts. Cancer. Res. 1996, 56: 2864-2871.
Zang, M.and C.H. Su. Ganoderma comphoratum, a new taxon in genus Ganoderma from Taiwan, China. Acta Bot. Yunnanica 1990, 12:395-396.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top