跳到主要內容

臺灣博碩士論文加值系統

(107.21.85.250) 您好!臺灣時間:2022/01/18 09:19
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:劉恆仲
研究生(外文):LIU HENG CHUNG
論文名稱:LL-37融合蛋白與化學合成LL-37以及生化合成LL-37之生產效益和抗菌功能的評估
論文名稱(外文):Evaluation of production performance and antimicrobial function of the LL-37 peptide as fusion protein and as chemically synthesized and biologically generated peptide
指導教授:陳震漢胡念台
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫微生物學研究所
學門:獸醫學門
學類:獸醫學類
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:107
中文關鍵詞:抗菌蛋白
外文關鍵詞:Antimicrobial peptide
相關次數:
  • 被引用被引用:0
  • 點閱點閱:980
  • 評分評分:
  • 下載下載:196
  • 收藏至我的研究室書目清單書目收藏:1
中文摘要
本實驗在評估化學合成的LL-37抗菌胜 (peptide) 和生化合成的LL-37融合蛋白的抗菌功能和生產效益。當人類上皮細胞受到微生物侵入引起局部性發炎時,會產生各種成份的胜,以消滅外來入侵的細菌。其中之一的基因是以豬的PR-39之cDNA作為探針,自人類骨髓cDNA基因庫分離到,此基因的蛋白產物由約170個氨基酸組成,它可被切割成二個片段,N端為cathelin propart,是一種cysteine protein inhibitor,C端為LL-37 antimicrobial peptide,以經過純化的LL-37,進行細菌抑制環分析(inhibition zone assay),顯示其具有抗菌性。它會形成a-helix的結構,且能溶解細菌細胞膜。利用E. coli量產LL-37抗菌胜,將此基因序列插入由125個氨基酸組成的 ketosteroid isomerase (KSI) 下游,及一段His-tag序列的上游。在E. coli表現的KSI/LL-37蛋白質形成的包涵體,自膠體切割回收和分離後,以細菌抑制環分析測試,顯示對革蘭氏陽性及陰性菌均有抗菌活性,但MIC比文獻報導略高。文獻指出,環境因子影響抗菌胜的結構和相異分子間作用的干擾,必須適當調控。接著我們也成功地利用大腸桿菌,表現出一段含三種功能區的LL-37/CBD/RGD融合蛋白質,它可能也受環境影響和融合蛋白的其他部份影響,抗菌效果需進一步研究。
Abstract
The aim of this study is to evaluate production performance and biological function of the chemically synthesized LL-37 peptide and the LL-37 as biologically generated fusion protein. Upon infection by microbes, human beings produce antimicrobial peptides as a defense mechanism. One of such genes was isolated from a cDNA library of human myeloid cells by probing with PR-39 cDNA of swine. Protein product of this gene is composed of 170 amino acid residues. It could be subdivided into two fragments. The N-terminal cathelin propart is a cysteine protein inhibitor and the C-terminal LL-37 peptide is an antimicrobial peptide. Inhibition zone assay indicated that purified LL-37 shows antimicrobial activity. Having been demonstrated to form a-helical structure, it could solubilize bacterial membrane. In order to overproduce LL-37 peptide in Escherichia coli, its coding sequence was inserted downstream of a 125 amino acid ketosteroid isomerase (KSI) gene and upstream of a His-tag sequence. The KSI/LL37 overproduced as inclusion bodies was purified from SDS-PAGE gel slice. Inhibition zone assay revealed that it has antimicrobial activity against both gram-positive and gram-negative bacteria, although its MIC appears higher than what has been reported. It has been reported that antimicrobial activity of peptide is influenced by various molecules in the environment and needs to be well controlled. In addition, we also produced in E. coli a tripartite chimeric protein LL-37/CBD/RGD. Its antimicrobial activity, probably also influenced by other components of the chimeric protein, needs to be re-evaluated.
[論文目次]
目 次
頁次
中文摘要………………………………………………………………1
英文摘要………………………………………………………………2
壹、 前 言…………………………………………………………..3
貳、 研究動機與目的…………………………….………………...5
參、 文獻探討………………………………………………………6
一、抗菌蛋白(胜)-LL-37的介紹..……………………………6
二、LL-37與抗生素的比較-抗菌機制、抗藥性、優缺點....….9
三、LL-37生產(化學與生化合成方法).………………………….14
四、融合蛋白質的可行性…………………………………………17
五、經濟效益評估…………………….…………………………..19
六、固定化……..……………………………………………….….21
七、研究目的……………………..…………………………….….22
肆、 材料與方法…………………………………………………..…23
一、LL-37化學合成法………………………………………...…..23
二、MIC (minimal inhibitory concentration) 抑制圈的測量…...25
三、LL-37基因片段……………………………………………….26
四、小量DNA質體之抽取………………………………………..26
五、大量DNA質體之抽取………………………………………..27
六、聚合酵素連鎖反應 ( Polymerase chain reaction )……………29
七、DNA的回收與萃取………….…………………………….….30
八、接合作用 (Ligation)………………………………………..….31
九、轉形作用 ( Transformation )………….……………………….32
十、定點突變法 ( Site-Directed Mutagenesis)………………..…...33
十一、重組蛋白的誘導表現…………………………………….....37
十二、不可溶 (KSI) 蛋白質表現系統的建立……………………38
十三、蛋白質膠體 ( SDS-PAGE ) 的電泳分析……………..……39
十四、蛋白質濃度的測定……………..………………………….…40
十五、KSI/LL-37的純化………………………………..……..……40
a. 親合性管柱分析法
b. 電淋洗 (Electro-elution)
十六、溴化氰 (CNBr) 切割生化合成的KSI/LL-37…………….42
十七、LL-37/CBD/RGD蛋白質的純化…………………………….43
十八、可溶 (Heat shock) 蛋白質表現系統的建立………………...44
伍、 結 果 …………………………………………………………...46
一. 化學合成的LL-37抗菌蛋白:
抗菌性分析……………………………………………….. 46
二. 生化合成的KSI/LL-37抗菌蛋白:
1. KSI/LL-37基因的構築……….………………………..46
2. KSI/LL-37的表現及分佈…………….………………..47
3. KSI/LL-37的純化……………………………………...48
a. 親合性色層分析管柱
b. 電淋洗 (Electron-elution)
4. 溴化氰 (cyanogen bromide) 切割含
KSI/LL-37包涵體………………………………………49
5. 胜色層分析管柱純化…..…………….………………50
6. PD-10色層分析管柱純化.………………………………51
7. 最小抑制濃度 (minimal inhibitory concentration)
試驗.………………………………………………….…51
三. 表現 LL-37/CBD/RGD融合蛋白質的基因構築………….51
四. LL-37/CBD/RGD融合蛋白質的誘導與
表現、純化與抗菌試驗………………………………………52
五. pCM7融合蛋白質的誘導表現與純化……………………….53
陸、 討 論 ……………………………………………………………..55
柒、 結 論………………………………………………………………..61
捌、 參考文獻……………………………………………………….…...63
玖、 附錄
培養液與試劑配方……………………………………………..…..70
圖 次………………………………………………………………..73
表 次………………………………………………………………105
圖次
化學合成儀…………………………………………………………….73
LL-37抗菌胜分析…………………………………………………..74
質譜儀鑑定化學合成的LL-37………………………………………..75
化學合成的LL-37的純度…………………………………………….76
化學合成的LL-37抑制大腸桿菌的測試結果…………………...…..77
化學合成的LL-37抑制枯草桿菌與沙門氏菌的測試結果……….…78
AlwNI與Pst I與限制酵素載體進行切割反應………………………79
pET-31b(+) 載體表現系統……..……………………………………..80
LL-37定序圖……………………………………………………….….81
插入pET-31b(+) 系統片段大小的結果………..…………………….82
pHC重組質體構築圖……………………………………………….…83
KSI/LL-37融合蛋白的誘導表現………………………………..…….84
KSI蛋白質的誘導表現………………………………………….…….85
KSI蛋白質的純化結果……………………………………….……….86
KSI/LL-37不可溶蛋白的誘導表現…………………………….……..87
含LL-37包涵體經鎳親合性管柱純化的結果…………..….………...88
電淋洗回收含LL-37包涵體的結果…………………………………..89
溴化氰 (CNBr) 切割LL-37包涵體的結果…………………………..90
化學合成LL-37經過胜色層管柱分析圖…………………………..91
生化合成的LL-37抑制革蘭氏陰性菌 (E. coli) 的測試結果……….92
生化合成的LL-37抑制革蘭氏陰性菌 (Salmonella) 的測試結果…..93
pHC-1載體構築圖……………………………………………………...94
pET-32a(+) 載體表現系統……………………………………..………95
LL-37/CBD/RGD定序圖…………………………………….….……...96
插入pET-32a(+) 系統片段大小的結果………………………….……97
pET表現系統……………………………………………………….…..98 LL-37/CBD/RGD融合蛋白質的誘導與表現……………………….....99
LL-37/CBD/RGD融合蛋白質的誘導與純化…………………….……100
LL-37/CBD/RGD融合蛋白質純化的結果……………………….……101
LL-37/CBD/RGD融合蛋白質抗菌測試……………………………….102
誘導與表現可溶性pCM7融合蛋白質的結果……………………...…103
pCM7融合蛋白質純化的結果…………………………………………104
表次
表一 Primer序列………………………………………………………105
表二 化學合成LL-37的抑菌測試結果………………………………106
表三 市面上胜價格……………………………………..……….….107
參考文獻
蔡文城。1996。微生物學。藝軒圖書出版社出版。9:170-195。
陳韻如。1997。開發CBD-RGD融合蛋白質應用於固定化細胞培養。碩士論文。國立中興大學獸醫所。台中市。中華民國。
陳靜美。1999。CBD融合蛋白質應用於檢驗試劑的開發。碩士論文。國立中興大學獸醫學系。台中市。中華民國。
孫崧桓 。1999。小口徑人工血管經表面改質後植覆內皮細胞之研究。碩士論文。國立中興大學化學工程研究所。台中市。中華民國。
謝淑枝。2000。Chitosan-Alginate-Hyaluronate複合物經RGD改質後的組織工程支架應用於軟骨再生之評估。碩士論文。國立中興大學化學工程研究所。台中市。中華民國。
江翰。2001。以RGD-EGFP融合蛋白作為細胞穿透現象之探討。碩士論文。國立中興大學獸醫微生物研究所。台中市。中華民國。
Agerberth, B., H. Gunne., J. Odeberg., P. Kogner., H. G. Boman, and G. H.
Gudmundsson. 1995. FALL-39, a putative human peptide antibiotic, is cysteine-
free and expressed in bone marrow and testis. Proc. Natl. Acad. Sci. USA. 92: 195—199.
Agerberth, B., J. Grunewald, and E. Castanos-Velez. 1999. Antibacterial components in bronchoalveolar lavage fluid from healthy individuals and sarcoidosis patients. Am J Respir Crit Care Med. 160: 283-290.
Andreas, P. 2002. How do bacteria resist human antimicrobial peptides ? Trends in Microbiology. Vol.10 No.4:179-186.
Assouline, Z., H. Shen., D. G. Kilburn, and R. A. Warren. 1993. Production and properties of a factor X-cellulose binding domain fusion protein. Protein Eng. 6: 787-792.
Athan, K. 1994. High-yield expression and purification of recombinant peptide. J Am. Chem. Soc. 116:4599-4607.
Bals, R., X. Wang., M. Zasloff, and J. M. Wilson. 1998. The peptide antibiotic LL-37/hCAP18 is expressed in epithelia of the human lung where it has broad antimicrobial activity at the airway surface. Proc. Natl. Acad. Sci. USA. 95: 9541—9546.
Bals, R., D. J. Weiner., A. D. Moscioni., R. L. Meegalla, and J. M. Wilson. 1999. Augmentation of Innate Host Defense by Expression of a Cathelicidin Antimicrobial Peptide. Infect. Immun. 67: 6084—6089.
Boman, H. G. 1995. Peptide antibiotics and their role in innate immunity.
Annu. Rev. Immunol. 13: 61—92.
Buchner, P. J. I., and U. Brinkmann. 1992. A method for increasing the yield of properly folded recombinant fusion proteins: single-chain immunotoxins from renaturation of bacterial inclusion bodies. Anal. Biochem. 205: 263-270.
Cole, A. M., J. Shi., A. Ceccarelli., Y. H. kim., A. Park, and T. Ganz. 2001. Inhibition of neutrophil elastase prevents cathelicidin activation and impairs clearance of bacteria from wounds. Blood. 97: 297-304.
Ellison-Ⅲ, R. T., and T. J. Giehl. 1991. Killing of gram-negative bacteria by and lysozyme. J. Clin. Invest. 88:1080-1091.
Frohm, M., B. Agerberth, G. Ahangari, M. Stahle-Backdahl, S. Liden, H. Wigzell, and G. H. Gudmundsson. 1997. The expression of the gene coding for the antibacterial peptide LL-37 is induced in human keratinocytes during inflammatory disorders. J. Biol. Chem. 272: 15258-15263.
Francis, M. J., G. Z. Hastings., B. E. Clarke., A. L. Brown., C. R. Beddell., D. J. Rowlands, and F. Brown. 1990. Neutralizing antibodies to all seven serotypes of foot-and-mouth disease virus elicited by synthetic peptides. Immunology. 69: 171-176.
Ganz, T., and R. I. Lehre. 1997. Antimicrobial peptides of leukocytes. Curr Opin Hematol. 4: 53-58.
Gebauer, F., J. C. D. L. Torre., I. Gomes., M. G. Mateu., H. Barahona., B. Tiraboschi., I. Bergmann., P. A. D. Mello, and E. Domingo. 1988. Rapid selection of genetic and antigenic variants of foot-and-mouth disease virus during persistence in cattle. J. Virol. 62: 2041-2049.
Greenwood, J. M., N. R. Gilkes., D. G. Kilburn., J. Miller., C. R, and R. A. J. Warren. 1989. Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose. FEBS Lett. 244:127-131.
Gudmundsson, G. H., B. Agerberth., J. Odeberg., T. Bergman., B. Olsson, and R. Salcedo. 1996. The human gene FALL39 and processing of the cathelin precursor to the antibacterial peptide LL-37 in granulocytes. Eur. J. Biochem. 238:325-332.
Hancock, R. E., and G. Diamond. 2000. The role of cationic antimicrobial peptides in innate host defences. Trends in Microbiology. 8: 402-410.
Hancock, R. E., and R. Lehrer. 1998. Cationic peptides: a new source of antibiotics. Trends in Biotech. 16: 82-88.
Harwig, S. S. L., A. J. Waring., H. J. Yang., Y. Cho., L. Tan, and R. I. Lehrer.
1996. Intramolecular disulfide bonds enhance the antimicrobial and lytic
activities of protegrins at physiological sodium chloride concentrations. Eur.
J. Biochem. 240: 352—357.
Haynie, S. L., G. A. Crum, and B. A. Doele. 1995. Antimicrobial activities of amphiphilic peptides covalently bonded to a water-insoluble resin. Antimicrob. Agents. Chemother. 39:301-307.
Hlavac, F., and E. Rouer. 1997. Expression of the protein-tyrosine kinase p56lck by the p vector yields a highly soluble protein recovered by mild sonication. Protein Expr. Purif. 11: 227-232.
Hsih, M. H., J. C. Kuo, and H. J. Tsai. 1997. Optimization of the solubilization and renaturation of fish growth hormone produced by Escherichia coli. App. Microbiol. Biotechnol. 48: 66-72.
Huttener, K. M., and C. L. Bevins. 1999. Antimicrobial peptides as mediators of epithelia host defense. Pediatric Res. 45: 785-794.
Jan, J., H. G. Gudmundur., E. R. Martin., D. B. Kurt, and A. Birgitta. 1998. Conformation-dependent Antibacterial Activity of the Naturally Occurring Human Peptide LL-37. J. Biol. Chem. 273:3718-3724.
Jensen, E. B., and S. Carlsen. 1990. Production of recombinant human growth hormone in E. coli: exprssion of different precursors and physiological effects of glucose, acetate, and salts. Biotech. Bioeng. 36: 1-11.
Kilburn, D. G., R. F. B. Turner., J. B. Coutinho., N. Din., N. R. Gilkes., J. M. Greenwood., J. B. Hobbs, and R. A. Warren. 1993. pp.45-54. Cellulose binding domains: applications in biotechnology, In: J. F. Kennedy, G. O. Phillips, P. A. Williams(eds.), CELLULOSICS: Materials for Selective Separations and other Technologies. Ellis-Horwood Press, [city where press located]. West Sussex, England.
Krebs, O., R. Ahl., O. C. Straub., O. Marquardt. 1992. Amino acid changes outside the G-H loop of capside protein VP1 of type O foot-and-mouth disase virus confer resistance to neutaliztion by antipeptide G-H serum. Vaccine. 11: 359-362.
LaVallie, E.R., E. A. DiBlasio., S. Kovacic., K. L. Grant., P. F. Schendel, and J. M. McCoy. 1993. A thioredoxin gene fusion expression system that circumvents inclusion body formation in the E. coli cytoplasm. Bio/Technology. 11(2): 187-193.
Lee, J. H., J. H. Kim., S. W. Hwang., W. J. Lee., H. K. Yoon., H. S. Lee, and S. S. Hong. 2000. High-level expression of antimicrobial peptide mediated by a fusion partner reinforcing formation of inclusion bodies. Biochem Biophys Res Commun. 277: 575-580.
Lehrer, R. I., A. Barton., K. A. Daher., S. S. L. Harwig., T. Ganz, and M. E.
Selsted. 1989. Interaction of human defensins with Escherichia coli: mechanism
of bactericidal activity. J. Clin. Invest. 84: 553—561.
Lilie, H., E. Schwarz, and R. Rudolph. 1998. Advances in refolding of proteins produced in E. coli. Curr. Opin. Biotechnol. 9: 497-501.
Matsuzaki, K. 1999. Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes. Biochim. Biophys. Acta. 1462: 1-10.
Merrifield, R. B. 1969. Solid-phase peptide synthesis. Adv Enzymol Relat Areas Mol Biol. 32:221-96.
Merrifield, R. B., E. L. Merrifield., P. Juvvadi., D. Andreu, and H. G. Boman. 1994. Design and systhesis of antimicrobial peptides. In “antimicrobial peptides”, 186: 5-26. Wiley, Chichester. (Ciba Foundation Symposium) Mor, A. 2000. Peptide-based antibiotics: a potential answer to raging antimicrobial resistance. Drug Develop Res. 50: 440-447.
Moks, T., L. Abrahmsen., B. Osterlof., S. Josephson., M. Ostling., S. O. Enfors., I. L. Persson., B. Nilsson, and M. Uhler. 1987. Large-scale affinity purification of human insulin-like growth factor I from culture medium of Escherichia coli. Bio/Technology. 5: 379-382.
Muhle, S. A., and J. P. Tam. 2001. Design of Gram-negative selective antimicrobial peptides. Biochemistry. 40: 5777-5785.
Nagaoka, I., S. Hirota., S. Yomogida., A. Ohwada, and M. Hirata. 2000. Synergistic actions of antibacterial neutrophil defensins and cathelicidins. Inflammation Research. 49: 73-79.
Ong, E., N. R. Gilkes., J. Miller., C. R., A. J. Warren, and D. G. Kilburn. 1991. Enzyme immobilization using a cellulose binding domain : properties of a β-glucosidase protein. Enzyme Microb. Technol. 13: 59-65.
Oswald, T., W. Wende., A. Pingoud, and U. Rinas. 1994. Comparison of N-terminal affinity fusion domains: effect on expression level and product heterogeneity of recombinant restriction endonuclease EcoRV. App. Microbiol. Biotechnol. 42: 73-77.
Pungor, E., N. B. Afeyan., N. F. Godon, and C. L. Cooney. 1987. Continuous affinity-recycle extraction: A novel protein separation technique. Bio/Technology. 5: 604-608.
Ramirez, C., J. Fung., R. C. J. Miller., R. Antony., J. Warren, and D. G. Kilburn. 1993. A bifunctional affinity linker to couple antibodies to cellulose. Bio / Technology. 11(13):1570-3.
Schein, C. H., and M. H. M. Noteborn. 1988. Formation of soluble recombinant proteins in Escherichia coli is favored by lower growth temperature. Bio/Technology. 6: 291-294.
Schonwetter, B. S., E. D. Stolzenberg, and M. A. Zasloff. 1995. Epithelial
antibiotics induced at sites of inflammation. Science. 267:1645—1648.
Studier, F. W., and B. A. Moffatt. 1986. Use of Bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned gens. J. Mol. Biol. 189:113-130.
Thomas, G., O. H. Sven., W. L. James., S. Ulrich, and W. Andre. 2001. Interaction of CAP18-Derived Peptides with Membranes Made from Endotoxins or Phospholipids. Biophys J. 80:2935—2945.
Turner, J ., Y. Cho., N. N. Dinh., A. J. Waring, and R. I. Lehrer. 1998. Activities of LL-37, a Cathelin-Associated Antimicrobial Peptide of Human Neutrophils. Antimicrob. Agents Chemother. 42:2206-2214.
Wierzba, A., U. Reichl., R. F. B. Turnner., R. A. J. Warren, and D. G. Kilburn. 1995. Production and Properties of a Bifunctional Fusion Protein that Mediates Attachment of Vero Cell to Cellulosic Matrices. Biotech. Bioeng. 47:147-154.
Wierzba, A., U. Reichl., R. F. B. Turnner., R. A. J. Warren, and D. G. Kilburn. 1995. Addachment of Mammailian Cell to a Recombiant Attachment Factor, CBD/RGD, Analyzed by Image Analysis. Biotech. Bioeng. 46:185-193.
Wu, M., E. Maier., R. Benz, and R. E. Hancock. 1999. Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemistry. 38: 7235-7242.
Yan, H., and R. E. Hancock. 2001. Synergistic interactions between mammalian antimicrobial defense peptides. Antimicrob. Agents Chemother. 45: 1558-1560.
Yang, L., T. M. Weiss., R. I. Lehrer, and H. W. Huang. 2000. Crystallization of antimicrobial pores in membranes: magainin and protegrin. Biophys J. 79 (4): 2002-2009.
Yee. L., and H. W. Blanch. 1992. Recombinant protein exprssion in high cell density fed-batch cultures of Escherichia coli. Biotechnol. 10: 1550-1556.
Yoon, S. K., W. K. Kang, and T. H. Park. 1994. Fed-batch operation of recombinant
E.coli containing trp promoter with controlled specific growth rate. Biotech. Bioeng. 43: 995-999.
Zanetti, M., R. Gennaro, and D. Romeo. 1995. Cathelicidins: a novel protein family with a common proregion and a variable C-terminal antimicrobial domain. FEBS. Lett. 374:1-5.
Ziv, O., C. L. Jeffrey., H. G. Gudmundur., A. Birgitta, and S. Yechiel. 1999. Structure and organization of the human antimicrobial peptide LL-37 in phospholipid membranes: relevance to the molecular basis for its non-cell-selective activity. Biochem. J. 341:501-513.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
1. 鄭富森(民88),目前教學評量之省思與改進之道,教師天地,99期,18─24頁。
2. 鄭富森、陳清義(民85),非紙筆測驗,測驗統計簡訊,9期,1-26頁。
3. 鄭富森(民87),淺談國中基本學力測驗之評鑑原則,測驗與輔導, 150期,3119-3125頁。
4. 陳文典(民87),實作評量在學力測驗之應用,測驗與輔導, 150期,3108-3111頁。
5. 莊明貞(民84),在國小課程改革的近程與發展─真實性評量,教師天地,79期,21─25頁。
6. 柯啟瑤(民89b),九年一貫課程研究與發展專刊5─基本能力的保障和評量,翰林文教雜誌,11期,1─2頁。
7. 柯啟瑤(民89a),九年一貫課程研究與發展專刊5─「自然與生活科技」的評量,翰林文教雜誌,11期,19─27頁。
8. 李虎雄、張敏雪(民88),由學力測驗方法談實作評量之功能,教師天地,99期,61─67頁。
9. 吳韻儀(民87),全球教改趨勢─教改大浪 襲捲全球,天下雜誌1988教育特刊─海闊天空Ⅱ跨世紀希望工程師,20─25頁。
10. 吳柏林、謝名娟(民88),自然潛能與數學生活化主義─談九年一貫數學課程教材教法,教育研究月刊,84期,94─頁。民90年3月10日,取自http://www.edujournal.com.tw/84/1.htm
11. 田耐青(民88),由多元智慧理論的觀點談教學評量:一些台灣的實例,教師天地,99期,32─37頁。
12. 簡茂發(民88),多元化評量之理念與方法,教師天地,99期,11─17頁。
13. 許學仁、何翠華(民89),國語科多元評量的實施,翰林文教雜誌,11期,8-11頁。