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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 許輔 | |
dc.contributor.author | Feng-Hsu Yang | en |
dc.contributor.author | 楊豐旭 | zh_TW |
dc.date.accessioned | 2021-06-13T01:08:58Z | - |
dc.date.available | 2007-07-27 | |
dc.date.copyright | 2007-07-27 | |
dc.date.issued | 2007 | |
dc.date.submitted | 2007-07-23 | |
dc.identifier.citation | 王志鵬。2001。 I. 枯草桿菌Bacillus subtilis DB104電轉形效率之增進 II. 設計、合成第一型抗凍蛋白基因及其於枯草桿菌、大腸桿菌中之表現。中興大學食品科學系研究所碩士論文。
杜元平。2003。 金針菇免疫調節蛋白FIP-fve抗腫瘤活性之探討。台灣大學園藝學研究所碩士論文。 周怡廷。2004。 以菌體表層展示技術表現重組之表層蛋白及融合蛋白於大腸桿菌、乳酸桿菌及乳酸鏈球菌。中興大學食品科學系研究所碩士論文。 唐嘉芳。2002。 乳酸菌對人體免疫力的影響與機制之探討。 台北醫學院保健營養學研究所碩士論文。 彭瑄第。2004。 乳酸菌發酵乳對降低過敏反應之影響。中興大學食品科學系研究所碩士論文。 廖啟成。1998。 乳酸菌之分類及應用。 食品工業。 30: 1-10。 潘子明。2005。 我國乳酸菌最近的研究趨勢暨通過健康食品認證之乳酸菌產品現況。農業生技產業季刊 3:19-27。 Aires, K. A., A. M. Cianciarullo, S. M. Carneiro, L. L. Villa, E. Boccardo, G. Perez-Martinez, I. Perez-Arellano, M. L. Oliveira, and P. L. Ho. 2006. Production of human papillomavirus type 16 L1 virus-like particles by recombinant Lactobacillus casei cells. Appl. Environ. Microbiol. 72:745-752. Arunachalam, K., H. S. Gill, and R. K. Chandra. 2000. Enhancement of natural immune function by dietary consumption of Bifidobacterium lactis (HN019). Eur. J. Clin. Nutr. 54:263-267. Banwart G. J. and V. N. Reinhold. 1989. Basic food microbiology. Van Nostrand Reinhold Publishers. Blatny, J. M., H. Ertesvag, I. F. Nes, and S. Valla. 2003. Heterologous gene expression in Lactococcus lactis; expression of the Azotobacter vinelandii algE6 gene product displaying mannuronan C-5 epimerase activity. FEMS Microbiol. Lett. 227:229-235. Brian J.B. and J. Warner. 2003. Genetics of lactic acid bacteria. Kluwer Academic/Plenum Publishers. Bruno-Barcena, J. M., J. M. Andrus, S. L. Libby, T. R. Klaenhammer, and H. M. Hassan. 2004. Expression of a heterologous manganese superoxide dismutase gene in intestinal lactobacilli provides protection against hydrogen peroxide toxicity. Appl. Environ. Microbiol. 70:4702-4710. Canzi, E., R. Zanchi, P. Comaschella, P. Cresci, , G. F. Grepp, C. Orpianesi, , M. Serrantoni and A. M. Ferrari. 2000. Modulation by lactic acid bacteria of the intestinal ecosystem and plasma cholesterol in rabbits fed a casein diet. Nutr. Res. 20: 1329–1340. Chaillou, S., P. H. Pouwels, and P. W. Postma. 1999. Transport of D-xylose in Lactobacillus pentosus, Lactobacillus casei, and Lactobacillus plantarum: evidence for a mechanism of facilitated diffusion via the phosphoenolpyruvate:mannose phosphotransferase system. J. Bacteriol. 181:4768-4773. Chiang, B. L., Y. H. Sheih, L. H. Wang, C. K. Liao, and H. S. Gill. 2000. Enhancing immunity by dietary consumption of a probiotic lactic acid bacterium (Bifidobacterium lactis HN019): optimization and definition of cellular immune responses. Eur. J. Clin. Nutr. 54:849-855. Clewell, D. B., Y. Yagi, G.. M. Dunny and Schultz S. K. 1974. Characterization of three plasmid deoxyribonucleic acid molecules in a strain of Streptococcus faecalis: identification of a plasmid determining erythromycin resistance. J. Bacteriol. 117:283–289. FAO/WHO. 2001. Report of a joint FAO/WHO expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Fuller, R. 1989. Probiotic in man and animals. J. Appl. Bacteriol. 66: 365-378. Gill, H. S., K. J. Rutherfurd, J. Prasad, and P. K. Gopal. 2000. Enhancement of natural and acquired immunity by Lactobacillus rhamnosus (HN001), Lactobacillus acidophilus (HN017) and Bifidobacterium lactis (HN019). Br. J. Nutr. 83:167-176. Gill, H. S., K. J. Rutherfurd, M. L. Cross, and P. K. Gopal. 2001. Enhancement of immunity in the elderly by dietary supplementation with the probiotic Bifidobacterium lactis HN019. Am. J. Clin. Nutr. 74:833-839. Gopal, P. K., J. Prasad, J. Smart, and H. S. Gill. 2001. In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli. Int. J. Food Microbiol. 67:207-16. Gosalbes, M. J., C. D. Esteban, J. L. Galan, and G. Perez-Martinez. 2000. Integrative food-grade expression system based on the lactose regulon of Lactobacillus casei. Appl. Environ. Microbiol. 66:4822-4828. Gosalbes, M. J., V. Monedero, C. A. Alpert, and G. Perez-Martinez. 1997. Establishing a model to study the regulation of the lactose operon in Lactobacillus casei. FEMS Microbiol. Lett. 148:83-89. Gosalbes, M. J., V. Monedero, and G. Perez-Martinez. 1999. Elements involved in catabolite repression and substrate induction of the lactose operon in Lactobacillus casei. J. Bacteriol. 181:3928-3934. Grangette, C., H. Muller-Alouf, P. Hols, D. Goudercourt, J. Delcour, M. Turneer, and A. Mercenier. 2004. Enhanced mucosal delivery of antigen with cell wall mutants of lactic acid bacteria. Infect. Immun. 72:2731-2737. Havenaar, R., J. H. J. Huis in’t Veld 1992: Probiotics: A general view. The lactic acid bacteria, the lactic acid bacteria in health and disease, Chapman & Hall, New York, published. 1:209–224. Ho, P. S., J. Kwang, and Y. K. Lee. 2005. Intragastric administration of Lactobacillus casei expressing transmissible gastroentritis coronavirus spike glycoprotein induced specific antibody production. Vaccine 23:1335-1342. Hose, H. and Sozzi, T. 1991. Biotechnology group meeting probiotics - factor fiction. J. Chem. Technol. Biotechnol. 51: 540-544. Josson, K., P.Soetaert, F. Michiels, H. Joos and J. Mahillon. 1990. Lactobacillus hilgardii plasmid pLAB1000 consists of two functional cassettes commonly found in other Gram-positive organisms. J. Bacteriol. 172: 3089–3099. Ko, J. L., C. I. Hsu, R. H. Lin, C. L. Kao and J. Y. Lin. 1995. A new fungal immunomodulatory protein, FIP-fve isolated from the edible mushroom, Flammulina Velutipes and its complete amino acid sequence. Eur. J. Biochem. 228: 244-249. Ko, J. L., S. J. Lin, C. I. Hsu, C. L. Kao and J. Y.Lin. 1997. Molecular cloning and expression of a fungal immunomodulatory protein, FIP-fve, from Flammulina Velutipes. J. Formos. Med. Assoc. 96:517-524 McKay, L. L. and Baldwin, A. K. 1990. Applications for biotechnology: present and future improvements in lactic acid bacteria. FEMS Microbio. Lett. 87:3–14. McCracken, A., M. S. Turner, P. Giffard, L. M. Hafner, and P. Timms. 2000. Analysis of promoter sequences from Lactobacillus and Lactococcus and their activity in several Lactobacillus species. Arch. Microbiol. 173:383-389. Natori, Y., Y. Kano, and F. Imamoto. 1988. Characterization and promoter selectivity of Lactobacillus acidophilus RNA polymerase. Biochimie. 70:1765-1774. Ogawa, M., K. Shimizu, K. Nomoto, M. Takahashi, M. Watanuki, R. Tanaka, T. Tanaka, T. Hamabata, S. Yamasaki, and Y. Takeda. 2001. Protective effect of Lactobacillus casei strain Shirota on Shiga toxin-producing Escherichia coli O157:H7 infection in infant rabbits. Infect Immun. 69:1101-1108. Oliveira, M. L., V. Monedero, E. N. Miyaji, L. C. Leite, P. L. Ho, and G. Perez-Martinez. 2003. Expression of Streptococcus pneumoniae antigens, PsaA (pneumococcal surface antigen A) and PspA (pneumococcal surface protein A) by Lactobacillus casei. FEMS Microbiol. Lett. 227:25-31. Parker, R. B. 1974. Probiotic, the other half of the antibiotic story. Anim. Nutr. Health. 29: 4-8. Perdigón, G.., C. M. Galdeano, J. C. Valdez, M. Medici. 2002. Interaction of lactic acid bacteria with the gut immune system. Eur. J. Clin. Nutr. 56: 21–26. Perez-Arellano, I., M. Zuniga, and G. Perez-Martinez. 2001. Construction of compatible wide-host-range shuttle vectors for lactic acid bacteria and Escherichia coli. Plasmid 46:106-116. Perez-Arellano, I. and G. Perez-Martinez. 2002. Structural features of the lac promoter affecting gusA expression in Lactobacillus casei. Curr. Microbiol. 45:191-196. Perez-Arellano, I. and G. Perez-Martinez. 2003. Optimization of the green fluorescent protein (GFP) expression from a lactose-inducible promoter in Lactobacillus casei. FEMS Microbiol. Lett. 222:123-127. Poo, H., H. M. Pyo, T. Y. Lee, S. W. Yoon, J. S. Lee, C. J. Kim, M. H. Sung, and S. H. Lee. 2006. Oral administration of human papillomavirus type 16 E7 displayed on Lactobacillus casei induces E7-specific antitumor effects in C57/BL6 mice. Int. J. Cancer 119:1702-9. Posno, M., R. J. Leer, N. van Luijk, M. J. van Giezen, P. T. Heuvelmans, B. C. Lokman, and P. H. Pouwels. 1991a. Incompatibility of Lactobacillus vectors with replicons derived from small cryptic Lactobacillus Plasmids and segregational instability of the introduced vectors. Appl, Environ, Microbiol, 57:1822-1828. Posno M., P. T. H. M. Heuvelmans, M. J. F. Van Giezen, B. C. Lokman, P. H. Pouwels and R. J. Leer. 1991b. Complementation of the inability of Lactobacillus strains to utilize D-xylose with D-xylose catabolism-encoding genes of Lactobacillus pentosus. Appl. Environ. Microbiol. 57:2764-2766. Pouwels, P. H. and J. A. Leunissen. 1994. Divergence in codon usage of Lactobacillus species. Nucleic Acids Res. 22:929-936. Repa, A., C. Grangette, C. Daniel, R. Hochreiter, K. Hoffmann-Sommergruber, J. Thalhamer, D. Kraft, H. Breiteneder, A. Mercenier, and U. Wiedermann. 2003. Mucosal co-application of lactic acid bacteria and allergen induces counter-regulatory immune responses in a murine model of birch pollen allergy. Vaccine 22:87-95. Robert, F. W. 2000. Molecular biology, 2th ed. WCB/McGraw-Hill, publisher. Roberts, T. A., F. A. Skinner. 1983. Food microbiology: advances and prospects. Academic Press Publishers. Salminen, S., A. von Wright, A. Ouwehand and M. Dekker. 2004. Lactic acid bacteria : microbiological and functional aspects. Marcel Dekker Publishers. Sheih, Y. H., B. L. Chiang, L. H. Wang, L. K Chuh and H. S. Gill. 2001: Systemic immunityenhancing effect in healthy subjects following dietary consumption of the lactic acid bacterium Lactobacillus rhamnosus HN001. J. Am. Coll. Nutr. 20: 149-156. Shu, Q. and H. S. Gill. 2002. Immune protection mediated by the probiotic Lactobacillus rhamnosus HN001 (DR20) against Escherichia coli O157:H7 infection in mice. FEMS Immunol. Med. Microbiol. 34:59-64. Singh, J., A. Rivenson, M. Tomita, S. Shimamura, N. Ishibashi, and B. S. Reddy. 1997. Bifidobacterium longum, a lactic acid-producing intestinal bacterium inhibits colon cancer and modulates the intermediate biomarkers of colon carcinogenesis. Carcinogenesis 18:833-841. Smacchi, E. and M. Gobbetti. 2000. Bioactive peptides in dairy products: synthesis and interaction with proteolytic enzymes. Food Microbiol.17:129-141. Szajewska, H., M. Kotowska, J. Z. Mrukowicz, M. Armanska, and W. Mikolajczyk. 2001. Efficacy of Lactobacillus GG in prevention of nosocomial diarrhea in infants. J. Pediatr. 138:361-365. Tannock, G.W. 1990. The microecology of lactobacilli inhabiting the gastrointestinal tract. Adv. Microb. Ecol. 11: 147-171. Toit, M. D., C. M. A. P. Franz, L. M. T. Dick, U. Schilinger, P. Haberer, B. Warlies, F. Ahrens and W. H. Holzapfel. 1998. Characterization and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol level, faeces pH and faeces moisture content, Int. J. Food Microbiol. 40:93–104. Tortuero, F., E. Fernández, P. Ruperéz, and M. Moreno. 1997. Raffinose and lactic acid bacteria influence caecal fermentation and serum cholesterol in rats. Nutr. Res. 17:41-49. Tsai, J. S., Y. S. Lin, B. S. Pan and T. J. Chen. 2006. Antihypertensive peptides and γ-aminobutyric acid from prozyme 6 facilitated lactic acid bacteria fermentation of soymilk. Process Biochem. 41.1282–1288 Tuomola, E., R. Crittenden, M. Playne, E. Isolauri, and S. Salminen. 2001. Quality assurance criteria for probiotic bacteria. Am. J. Clin. Nutr. 73: 393–398 Vinderola, C. G., M. Medici and G. Perdigón. 2004. Relationship between interaction sites in the gut, hydrophobicity, mucosal immunomodulating capacities and cell wall protein profiles in indigenous and exogenous bacteria. J. Appl. Microbiol. 96: 230–243. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/29509 | - |
dc.description.abstract | 本研究將構築乳酸菌 (LAB) 異蛋白表現系統以獲得 fve 重組蛋白表現菌株,並探討 LAB 表現 fve 重組蛋白之免疫調節活性。
本研究首先構築 LAB 表現異蛋白之載體系統,以穿梭載體 pLP3537 作為骨架,再以 Lactobacillus casei ATCC 393 之 plac 啟動子作為啟動表現元件,搭配大腸桿菌商業載體 pQE30 之 lamda 終止子,建構表現載體 pLP-LT1;分別將 fve 蛋白以及 hisfve 蛋白之表現基因與 pLP-LT1 黏合,獲得表現載體 pLP-LFT1 及 pLP-LHFT1。成功獲得 LAB 重組菌株後,以乳糖培養基進行菌株培養及蛋白表現之誘導。由 SDS-PAGE 電泳結果顯示乳酸菌生產之重組 fve 及 hisfve 表現量極低,但 LAB/hisfve 總蛋白經由 FPLC 系統純化濃縮後,可獲得分子量約 18 kDa 之產物,並以 Anti-His tag 抗體可辨認到此蛋白。 以 BALB/c 小鼠進行免疫活性分析,非特異性免疫反應方面,結果顯示餵食 LAB、LAB/hisfve 及 LAB/fve 對小鼠腹腔巨噬細胞分泌 TNF-alpha 及 NO 以及刺激脾細胞增生及分泌 IFN-gamma 之活化均不顯著 (P>0.05)。在 OVA 特異性免疫反應方面,LAB、LAB/hisfve 及 LAB/fve 均可顯著提升小鼠脾細胞 OVA 特異性之細胞增生以及 OVA 特異性 IFN-gamma 產量,以及血清中分泌 OVA 特異性IgG (P<0.05)。然而 LAB/hisfve 及 LAB/fve 組與LAB組之間並無差異 (P>0.05) 。 本研究成功構築乳酸菌表現載體 pLP-LT1,獲得可表現 fve 蛋白之重組乳酸菌株,結果顯示此表現平台具有實際應用於食品工業之潛力。 | zh_TW |
dc.description.abstract | The objectives of this study were to construct a heteroprotein expression system of lactic acid bacteria (LAB), to express an immunomodulatory protein from Flammulina velutipes (fve) in recombinant LAB strain, and to study the immunoregulatory activity of fve-expressing LAB strains in vivo.
In this study, the LAB expression vector pLP-LT1 was constructed based on shuttle vector pLP3537 as a backbone with a plac promoter from Lactobacillus casei ATCC 393 and a lamda terminator from E.coli vector pQE30. The genes encoding hisfve and fve were cloned to generate the expression vectors pLP-LHFT1 and pLP-LFT1, respectively. After transformation, these two recombinant LAB stains were established and their expression of proteins was produced under lactose- induction. SDS-PAGE analysis showed that the expression levels of hisfve and fve proteins by LAB were too low for detection, although hisfve protein could be blotted and recognized by His-tag antibody after FPLC purification. Different LAB samples (LAB, LAB/hisfve, and LAB/fve) were orally fed (5x109 cells/day) to BALB/c mice for 30 days and their immunomodulatory activities were further evaluated. Results indicated that oral administration of LAB, LAB/fve or LAB/hisfve showed neither significant stimulation on TNF-alpha and NO production by mouse peritoneal macrophage (P>0.05) nor significant stimulation on cell proliferation and IFN-gamma secretion by spleen cells (P>0.05). Furthermore, oral administrations of LAB, LAB/fve or LAB/hisfve were effective in increasing OVA-specific cell proliferation and OVA-specific IFN-alpha secretion by spleen cells. The OVA-specific IgG production in mouse sera was also enhanced significantly (P<0.05). Unfortunately, the low fve-expressing capability of recombinant LAB could lead to insignificant immune activation between LAB, LAB/hisfve and LAB/fve treatments in vivo. Taking together, a LAB shuttle/expression vector pLP-LT1 was constructed and fve-expressing LAB strains were established in this study, which showed potentials for further utilization in food applications. | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T01:08:58Z (GMT). No. of bitstreams: 1 ntu-96-R94628211-1.pdf: 2056781 bytes, checksum: a5c630ded10cb601df0450fb259cfebd (MD5) Previous issue date: 2007 | en |
dc.description.tableofcontents | 目錄 i
圖表目錄 iv 中文摘要 1 英文摘要 3 第一章 研究背景 5 一、 前言 5 二、 益生菌與乳酸菌 6 三、 乳酸菌於食品之應用 7 四、 乳酸菌的基因工程 10 五、 構築乳酸菌選殖載體 13 六、 乳酸菌之保健功效 15 七、 乳酸菌與免疫機制調控 18 八、 金針菇免疫調節蛋白之相關研究 20 九、 研究目的與動機 22 第二章 材料與方法 23 第一節 菌株與質體 23 第二節 乳酸菌表現載體之構築 23 一、質體 DNA 分離 24 二、DNA 之剪切、回收及黏合 26 三、質體 DNA 之轉形法 28 四、表現元件基因之擴增 30 第三節 金針菇免疫調節蛋白於乳酸菌表現與偵測 33 一、乳酸菌之培養與菌體破碎 33 二、乳酸菌表現蛋白之純化 34 三、SDS-聚丙烯醯胺電泳分析 35 四、西方轉漬分析 37 第四節 體內非特異性免疫調節活性試驗 39 一、餵食乳酸菌之試驗設計 40 二、對腹腔巨噬細胞與脾細胞活化之影響 40 第五節 體內特異性免疫調節活性試驗 47 一、餵食乳酸菌之試驗設計 48 二、對小鼠脾細胞之影響 49 三、對小鼠血清中 OVA 特異性 IgG 之影響 52 第六節 統計分析 53 第三章 結果 55 第一節 於乳酸菌表現 fve 蛋白 55 一、乳酸菌表現載體之構築 55 二、fve 蛋白於乳酸菌之表現與偵測 59 第二節 fve 蛋白於乳酸菌表現對小鼠非特異性免疫調節活性 60 一、 對小鼠腹腔巨噬細胞之活化作用 60 二、 對小鼠脾貼附細胞之活化作用 62 第三節 fve 蛋白於乳酸菌表現對小鼠 OVA 特異性免疫調節活性 64 一、對小鼠脾細胞之 OVA 特異性活化作用 64 二、對小鼠血清中 OVA 特異性 IgG 之影響 65 第四章 討論 67 第五章 結論 73 參考文獻 74 圖與表 81 | |
dc.language.iso | zh-TW | |
dc.title | 金針菇免疫調節蛋白於乳酸菌表現及其免疫調節活性 | zh_TW |
dc.title | Studies on the Expression of Fungal Immunomodulatory Protein from Flammulina velutipes in Lactic Acid Bacteria and its Immunomodulatory Activity | en |
dc.type | Thesis | |
dc.date.schoolyear | 95-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 簡伯容 | |
dc.contributor.oralexamcommittee | 何國傑,柯俊良,繆希椿 | |
dc.subject.keyword | 乳酸菌,金針菇免疫調節蛋白,表現系統,免疫調節, | zh_TW |
dc.subject.keyword | lactic acid bacteria,fve,expression system,immunomodulatory, | en |
dc.relation.page | 107 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2007-07-23 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 園藝學研究所 | zh_TW |
顯示於系所單位: | 園藝暨景觀學系 |
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