跳到主要內容

臺灣博碩士論文加值系統

(216.73.217.144) 您好!臺灣時間:2026/04/25 13:50
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:闕立楓
研究生(外文):Li-Feng Chir
論文名稱:斑馬魚之卵黃蛋白辨識紫外光傷害DNA之研究
論文名稱(外文):Low-molecular-weight yolk proteins are components of a DNA damage binding activity highly expressed in zebrafish (danio rerio) embryos
指導教授:許濤許濤引用關係
學位類別:碩士
校院名稱:國立臺灣海洋大學
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:2004
畢業學年度:92
語文別:中文
論文頁數:59
中文關鍵詞:斑馬魚卵黃蛋白
相關次數:
  • 被引用被引用:0
  • 點閱點閱:284
  • 評分評分:
  • 下載下載:15
  • 收藏至我的研究室書目清單書目收藏:0
根據先前本實驗室之研究成果,已知在凝膠阻抗實驗中,斑馬魚之12 hr胚胎萃取液對UV-damaged-DNA具有高度的辨識能力(26)。我們將膠片上辨識蛋白與6-4 photoproduct(6-4PP)形成之複合體挖下,並取其中分子量為30 kDa之辨識蛋白作質譜分析,發覺其序列與斑馬魚之卵黃蛋白前軀物質vitellogenin I (zfVg 1)相似(已知Vg在胚胎發育時期會分解為卵黃蛋白)(32)。為研究Vg與卵黃蛋白對損傷DNA辨識之重要性,於凝膠阻抗實驗之反應物中加入抗鯉魚Vg血清,發現能抑制12 hr斑馬魚胚胎萃取液對6-4PPs之辨識能力。而當凝膠阻抗實驗反應物加入抗斑馬魚之Vg單株抗體濃度達6.4至18.8 μg/ml時,12 hr斑馬魚胚胎萃取液對6-4PPs之辨識能力僅為未加抗體實驗組別強度之80%至40%。然而加入濃度為18.8 μg/ml 之HSP70單株抗體並無抑制12 hr粹取液對6-4PPs之辨識能力。UV交錯連結實驗(UV-crosslinking)之反應物中加入斑馬魚之Vg單株抗體亦會抑制辨識蛋白對於6-4PPs之辨識能力。在斑馬魚胚胎萃取液中能被Vg抗體辨識之卵黃蛋白分子在不同發育時期(12, 48, 96 hr)有不同程度之表現。經過親和性吸附與西方點墨法分析,發現12 hr胚胎萃取液中分子量為30 kDa及35 kDa之卵黃蛋白對6-4PPs辨識能力有明顯的專一性,然而對UV-damaged-DNA辨識力甚低之96 hr幼魚萃取液中,上述30 kDa及35 kDa之卵黃蛋白分子含量甚低且其對6-4PPs之辨識能力幾乎無法偵測。透過UV交錯連結實驗以及西方點墨法實驗發現12 hr斑馬魚胚胎萃取液中之低分子量(30,35 kDa)卵黃蛋白並無直接參與UV-damaged-DNA之辨識。根據實驗結果我們推論卵黃蛋白除了提供斑馬魚胚胎發育時期所需之營養素(30)之外,低分子量之卵黃蛋白可能與其他蛋白結合參與辨識早期胚胎發育中對類似紫外光傷害造成之DNA扭曲結構。
名詞縮寫………………………………………p.4
英文摘要………………………………………p.5
中文摘要………………………………………p.7
前言……………………………………………p.9
實驗材料與方法………………………………p.14
實驗結果………………………………………p.30
討論……………………………………………p.35
參考文獻………………………………………p.41
附圖……………………………………………p.46
(1) Sinha RP, Hader DP.2002. UV-induced DNA damage and repair: a review. Photochem. Photobiol. 1(4):225-36
(2) Mitchell DL,Nairn RS.1989. The biology of the (6-4)photoproduct. Photochem. Photobiol. 49(6):805-19.
(3) Mizukoshi T, Fujiwara Y, Iwai S.1999. DNA structures recognized by the human UV-DDB protein. Nucleic Acids Symp. Ser. 42:265-6.
(4) Costa RM, Chiganças V, Galhardo Rda S, Carvalho H, Menck CF .2003. The eukaryotic nucleotide excision repair pathway . Biochimie 85 1083–99
(5) Kim ST, Malhotra K, Ryo H, Sancar A, Todo T. 1996. Purification and characterization of Drosophila melanogaster photolyase. Mutat. Res. 363(2):97-104.
(6) Otrin VR, McLenigan M, Takao M, Levine AS, Protic M. 1997. Translocation of a UV-damaged DNA binding protein into a tight association with chromatin after treatment of mammalian cells with UV light. J. Cell. Sci. 110(10):1159-68
(7) Rybanska I, Pirsel M.2003.Involvement of the nucleotide excision repair proteins in the removal of oxidative DNA base damage in mammalian cells.Neoplasma.50(6):389-95
(8) Thoma F. 1999.Light and dark in chromatin repair :repair of UV-induced DNA lesions by photolyase and nucleotide excision repair .EMBO J. 18(23):6585-98.
(9) Carell T, Burgdorf LT, Kundu LM, Cichon M. 2001. The mechanism of action of DNA photolyase. Curr. Opin. Chem. Biol. 5(5):491-8.
(10) Hwang BJ., Chu G. 1999. Purification and characterization of a human protein that binds to damaged DNA.
Biochemistry 32: 1657–66.
(11)Shiyanov P., Hayes SA., Donepudi M., Nichols AF., Linn S.,Slagle BL., Raychaudhuri P. 1999. The naturally occurring mutants of DDB are impaird in stimulating nuclear import of the p125 subunit and E2F1-avtivated transcription. Mol .Cell. Biol.19:4935-43.
(12)Otrin VR, McLenigan M, Takao M, Levine AS, Protic M. 1997. Translocation of a UV-damaged DNA binding protein into a tight association with chromatin after treatment of mammalian cells with UV light. J. Cell. Sci. 110(10):1159-68
(13) Fitch ME., Nakajima S., Yasui A., Ford J.M. 2003. In vivo recruitment of XPC to UV-induced cyclobutane pyrimidine dimers by the DDB2 gene product, J. Biol. Chem. 278:46906–10.
(14) Hayes S., Shiyanov X., Chen P., Raychaudhuri P. 1998. DDB, a putative DNA repaire protein ,can function as a transcriptional partner of E2F1. Mol. Cell. Biol. 18:240-49
(15)Vichi P, Coin F, Renaud JP, Vermeulen W, Hoeilmakers JH, Moras D, Egly JM. 1997.Cisplatin- and UV-damaged DNA lure the basal transcription factor TFIID/TBP. EMBO J. 16(24):7444-56
(16) Wood RD, Araujo SJ, Ariza RR, Batty DP, Biggerstaff M, Evans E,Gaillard PH, Gunz D, Koberle B, Kuraoka I, Moggs JG, Sandall JK, Shivji MK. 2000. DNA damage recognition and nucleotide excision repair in mammalian cells. Cold Spring Harb. Symp. Quant. Biol. 65:173-82.
(17) Hanawalt PC. 2002. Subpathways of nucleotide excision repair and their regulation. Oncogene 21: 8949–56.
(18) Seki N, Hayashi A, Hattori A, Kozuma S, Sasaki M, Suzuki Y,Sugano S, Muramatsu M, Saito T. 1999. cDNA clonging, tissue expression, and chromosomal assignment of a mousegene, encoding a 127 kDa UV-damaged DNA binding protein which is defective in XPE cell. DNA Res. 6(5):319-22.
(19) Lehman AR. 2003. DNA repair-deficient diseases, xeroderma pigmentosum,Cockayne’s syndrome and trichothiodystrophy, Biochimie. 84 :1044-52
(20) Tanaka K., Miura N., Satokata I., Miyamoto I., MC. Yoshida, Satoh Y., Kondo S., Yasui A., Okayama H., Okada Y. 1990. Analysis of a human DNA excision repair gene involved in group A xeroderma pigmentosum and containing a zinc-finger domain. Nature 348: 73–76.
(21) Wakasugi M., Sancar A. 1998. Assembly,subunit composition, and footprint of human DNA repair excision nuclease.
Proc. Natl. Acad. Sci.95:6669-74.
(22) Yang ZG., Liu Y., Mao LY., Zhang JT., Zhou Y. 2002. Dimerization of human XPA and formation of XPA2–RPA protein complex. Biochemistry 41: 13012–20.
(23) Kimmel CB., Ballard WW., Kimmel SR., Ullmann B., Schilling, TF. 1995. Stages of embryonic development of the zebrafish.
Dev. Dyn.203:253-310.
(24) Pasheva EA., Pashev IG., Favre A. 1998. Preferential binding of high mobility group 1 protein to UV-damaged DNA.
J. Biol.Chem.273:24730-6.
(25)Henrik H, Lene A . 2001. Development of an ELISA for vitellogenin in whole body homogenate of zebrafish.
Comp. Biochem. Physiol. C 130: 119-31.
(26) Hsu T, Cheng CS, Shih CY. Yeh FL. 2002. Detection and partial characterization of a UV-damaged-DNA binding activity highly expressed in zebrafish(Danio rerio) embryos. Fish Physiol. Biochem. 25: 41-51.
(27) Prisco M, Valiante S, Romano M.2004.
Ovarian Follicle Cells in Torpedo marmorata Synthesize Vitellogenin. Molecular Reproduction and Development
67:424-29.
(28)Kime DE. 1998. Disruption of liver function.In:Endocrine Disruption in Fish. Comp. Biochem. Physiol. C 124: 11-21
(29) Romano M, Limatola E. 2000. Oocyte plasma membrane proteins and the appearance of vitellogenin binding protein during oocyte growth in the lizard Podarcis sicula. Gen. Comp. Endocrinol. 118:383-92.
(30) Falchuk KH. 1998. The molecular basis for the role of zinc in
developmental biology. Mol. Cell Biochem. 188: 41–48.
(31) Wang H, Yan T, Tan JTT, Gong Z. 2000. A zebrafish vitellogenin gene (vg3) encodes a novel vitellogenin without a phosvitin domain and may represent a primitive vertebrate vitellogenin gene.
Gene 256: 303-10.
(32) Naoshi H, Norimasa I, Haruhisa F,Toshiaki F,Craig V. 2002. Identification and Characterization of Proteases Involved in Specific Proteolysis of Vitellogenin and Yolk Proteins in Salmonids.
Journal of Expepimental Zoology. 292:11–25
(33) Yokota Y, Unuma T, Moriyama A, Yamano K. 2003. Cleavage site of a major yolk protein(MYP) determined by cDNA isolation and amino acid sequencing, Hemicentrotus pulcherrimus.
Comp. Biochem. Physiol. B 135:71-81.
(34) Sappington TW, Raikhel AS. 1998. Mini review. Molecular characteristics of insect vitellogenins and vitellogenin receptors. Insect Biochem. Mol. Biol. 28:277– 300.
(35) You JS, Wang M, Lee SH. 2003. Biochemical analysis of the damage recognition process in nucleotide excision repair. J. Biol. Chem. 278: 7476-85.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top