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研究生:陳玲華
研究生(外文):Ling-Hua Chen
論文名稱:人類多瘤性病毒,JCV,主要殼體蛋白VP1功能性之分子與免疫反應研究
論文名稱(外文):Molecular and Immunological Analysis of the Human Polyomavirus,JC Virus,Major Capsid Protein VP1
指導教授:張德卿林培正林培正引用關係
指導教授(外文):Deching ChangPei-Cheng Lin
學位類別:碩士
校院名稱:中山醫學院
系所名稱:免疫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:54
中文關鍵詞:JC 病毒病毒殼體
外文關鍵詞:JC virusVP1 polypeptidecapsid assembly
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中文摘要
人類多瘤性病毒(JC virus)是乳多瘤病毒科(Papovaviridae)其中一
屬。病毒殼體(capsid)的組裝機制是由5 個主要殼體蛋白VP1 構成次殼體
(capsomere )、72 個次殼體組成病毒殼體,本研究主題探討JC 病毒主
要殼體蛋白VP1 組裝成病毒殼體的最低需求,分別於VP1 月生月太鏈
(polypeptide)的N 端刪除掉12(△N12)及19(△N19)個胺基酸以及C 端刪
除掉16(△C16)、17(△C17)、31(△C31)個胺基酸,於原核(prokaryotic)
細胞中表達蛋白質。經10%-30%及5%-20%蔗糖梯度離心和電子顯微鏡
觀察病毒的殼體結構是否受到影響。結果顯示△N12 、△C16 可自行組裝
成病毒殼體與WT VP1 相似;△N19 、△C17 、△C31 無法組裝成病毒殼
體以次殼體型態存在。經由這些結果推測出JC 病毒主要殼體蛋白VP1 月生
月太鏈形成病毒殼體的最低需求為N 端第19 個胺基酸以及C 端第17 個胺
基酸之間的區域。多瘤性病毒JC 病毒、SV40 病毒和Py 病毒這3 種多瘤
性病毒DNA 序列分析比對約有50%以上的相似度,但是宿主範圍有極大
的差異,且位於病毒殼體表面區域的BC 、DE 、HI loop 是序列分析比對
差異最大的地方。經由免疫反應分析多瘤性病毒殼體,由結果推測出病毒
殼體表面區域於病毒選擇宿主時是主要決定區域。
Abstract
Human JC virus (JCV) belongs to the family of polyomaviridae. The
viral capsid is composed of 72 capsomeres. Five VP1 molecules make
up a capsomere structure. To investigate the minimal sequences on JCV
VP1 polypeptide required for capsid assembly, the first twelve (△N12)
and nineteen (△N19) amino acids at the N-terminus and the last sixteen
(△C16), seventeen (△C17) and thirty-one (△C31) amino acids at the
C-terminus of VP1 were truncated and expressed in E. coli. The VP1
proteins of △N12 and △C16 were able to self-assemble into a
virus-like particle similar to that of wild type (WT) VP1. However, the
mutant proteins of △N19, △C17 and △C31 formed a pentameric
capsomere structure as demonstrated by a 10-30% sucrose gradient
centrifugation and 5-20% sucrose gradient centrifugation and electron
microscopy. These results suggest that the first nineteen and the last
seventeen residues are the minimal sequences on VP1 polypeptide for
JCV capsid assembly. Although DNA sequences of JCV,SV40 and Py
are highly homologous,their host ranges are different. The results of
immunological analysis showed that the surface domains,BC,DE and HI
loops of the viruses may play an important role in host determination.
目 錄
頁次
英文摘要 ----------------------------------------------------------------------------3
中文摘要 --------------------------------------------------------------------------- 4
前言
人類多瘤性病毒 JC 病毒背景介紹-------------------------------------------5
本研究相關資料簡介-------------------------------------------------------------9
材料與方法-------------------------------------------------------------------------11
結果----------------------------------------------------------------------------------21
討論----------------------------------------------------------------------------------26
圖表----------------------------------------------------------------------------------29
參考文獻----------------------------------------------------------------------------48
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