DC 欄位 | 值 | 語言 |
dc.contributor.author | 賴秀穗 | zh_TW |
dc.creator | 賴秀穗 | zh_TW |
dc.date | 2000 | zh_TW |
dc.date.accessioned | 2006-07-26T05:37:04Z | - |
dc.date.accessioned | 2018-07-09T06:15:40Z | - |
dc.date.available | 2006-07-26T05:37:04Z | - |
dc.date.available | 2018-07-09T06:15:40Z | - |
dc.date.issued | 2000 | - |
dc.identifier | 892313B002069 | zh_TW |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/28680 | - |
dc.description.abstract | 豬繁殖與呼吸道症候群(Porcine reproductive
and respiratory syndrome, PRRS) 與假性狂犬
病(Peudorabies, PR)為豬重要的傳染病。為了
研究可否利用減毒假性狂犬病病毒做為載體來攜
帶一段PRRS 病毒基因,以便構築雙價疫苗,做為
防疫該兩大疾病之用. 本實驗將PRRS 病毒的GP3
醣蛋白基因(即ORF3 基因)植入取代PR 病毒醣蛋
白基因內,以構築PRRS/PR 複合病毒(chimeric
virus)。本實驗首先利用反轉錄聚合脢鏈反應增
幅出PRRS 病毒的GP3 醣蛋白基因片段,再將之植
入pGEX-2T 原核表現載體, 並進行大量表現獲得
分子量約30 kDa 的GST-GP3 融合重組蛋白,然後
利用此重組蛋白製備單株抗體做為篩選和確認
PRRS/PR 複合病毒之用。經過單株抗體的融合製
備,順利獲得抗GP3 重組蛋白單株抗體,該單株抗
體具有辨識PRRS 病毒GP3 醣蛋白的能力;但不具
中和PRRS 病毒功能。另一方面,同樣利用反轉錄
聚合脢鏈反應的選殖方式,將PRRS 病毒的GP3 醣
蛋白基因,選殖入含PR 病毒gG 基因片段的pGX
載體內,再將含GP3 基因的pJ3X 重組質體與減毒
的PRV-gE--TK-病毒完整DNA 共同感染RK-13 細胞
進行同源重組(Homologous recombination)後,
成功構築出PRRS/PR 重組複合病毒(J3X),經由
南方轉漬試驗、西方免疫轉漬試驗、免疫螢光染色
試驗及免疫過氧化脢染色試驗的驗證得知J3X 複
合病毒基因體內確實嵌有PRRS 病毒的GP3 醣蛋白
基因,並可表現出GP3 重組醣蛋白。J3X 複合病毒
在細胞株所形成的病毒斑,比PR 病毒強毒株的病
毒斑小,未來將進行該複合病毒的抗原性試驗。 | zh_TW |
dc.description.abstract | Porcine reproductive and respiratory
syndrome (PRRS) and Pseudorabies (PR) are two
impor tant infectious diseases of swine. In order
to construct a chimer ic virus, PR virus was used as
a vector to car ry a foreign virus gene fragment
ORF 3 of PRRS. The total fragment of ORF3
gene was amplified by Rever se-Transcr iption
Polymerase Chain Reaction (RT-PCR). The
resulting cDNA was inser ted into prokaryotic
expression vector (pGEX-2T) system. GST-GP3
(gluthione-S-Transferase-glycoprotein5) expressed
fusion proteins having molecular weight about 30
kDa was obtained. The fusion proteins were used
as antigens to prepare monoclonal antibodies
(Mabs), and a hybr idoma capabling of secreting
the Mabs against the GP3 fusion proteins was
successfully produced. The Mabs against GP3
fusion proteins was used for screening and
confirming PRRS/PR chimer ic virus car rying the
ORF 3 gene expressed proteins. The Mabs can
identify native GP3 proteins of PRRS virus, but
not to neutralize PRRS virus. . Fur thermore, the
RT-PCR products GP3 gene fragments were
cloned into the pGX vector contained the gG gene
fragment of PR virus. Then, the constructed pJ3X
plasmid DNA contained gG gene fragment of PR
virus and GP3 gene fragment of PRRS virus and
DNA of the attenuated PRV-gE--TK- virus were
co-transfected into RK-13 cells. Following
homologous recombination in the cells, a PRRS/PR
chimer ic virus (designated as J3X chimer ic virus)
car r ied GP3 gene fragment in the genome of PR
virus was successfully constructed. GP3
recombinant proteins of PRRS/PR chimer ic virus
were detected by Southern blotting, Western
blotting, Immunofluorescent assay and
Immuno-peroxidase staining. Plaques produced
by J3X chimer ic virus in RK-13 cell lines were
much smaller than those produced by wild-type PR
virus. The chimer ic virus J3X will be fur ther
investigated on its immunogenicity in pigs. | en |
dc.format | application/pdf | zh_TW |
dc.format.extent | 32630 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | zh-TW | zh_TW |
dc.language.iso | zh_TW | - |
dc.publisher | 臺北市:國立臺灣大學獸醫學系暨研究所 | zh_TW |
dc.rights | 國立臺灣大學獸醫學系暨研究所 | zh_TW |
dc.subject.classification | [SDGs]SDG3 | - |
dc.title | 行政院國家科學委員會專題研究計畫成果報告:豬繁殖與呼吸道症候群及假性犬病複合疫苗之開發 | zh_TW |
dc.type | report | en |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/28680/1/892313B002069.pdf | - |
dc.coverage | 計畫年度:89
第一期;起迄日期:1999-08-01/2000-07-31 | zh_TW |
item.fulltext | with fulltext | - |
item.languageiso639-1 | zh_TW | - |
item.openairecristype | http://purl.org/coar/resource_type/c_93fc | - |
item.cerifentitytype | Publications | - |
item.openairetype | report | - |
item.grantfulltext | open | - |
顯示於: | 獸醫學系
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