DC 欄位 | 值 | 語言 |
dc.contributor | 國立臺灣大學醫學院外科 | zh_TW |
dc.contributor.author | 李伯皇 | zh_TW |
dc.creator | 李伯皇 | zh_TW |
dc.date | 2003-07-31 | - |
dc.date.accessioned | 2006-07-26T03:41:31Z | - |
dc.date.accessioned | 2018-07-11T09:26:45Z | - |
dc.date.available | 2006-07-26T03:41:31Z | - |
dc.date.available | 2018-07-11T09:26:45Z | - |
dc.date.issued | 2003-07-31 | - |
dc.identifier | 912314B002164 | zh_TW |
dc.identifier.uri | http://ntur.lib.ntu.edu.tw//handle/246246/24489 | - |
dc.description.abstract | 已發表的報告中已指出,缺氧後的肝
臟會有促進肝再生的現象,而且兩者成正
相關的關係;另有報告發現在肝臟切除手
術前先給予刺激(如剖腹探查)同樣的可
以增進肝臟的再生,但確切的機制及基因
被誘發表現或抑制表現的程度其增減上並
沒有研究報告得說明之。
本研究將以大鼠肝臟缺氧後(各處理
為:缺血零分鐘,缺血三十分鐘,缺血六
十分鐘)再行切除的模式,收集術後6 小
時及24 小時的肝臟檢體研究缺氧對肝臟
切除後肝再生的影響,並將收集的檢體儲
存於-70℃。實驗時將檢體取出部份以液態
氮及研缽研磨後萃取mRNA反轉錄成第一
股cDNA 後再合成第二股cDNA,合成的
cDNA 以兩種不同切位的限制脢切割酵素
切割後,接上特殊人為設計的轉接器即形
成兩端具已知序列的cDNA 片段。
進行AFLP 分析時則以外加2~3 個氨
基酸的引子數對來進行聚合脢放大反應放
大cDNA 片段以供定序電泳分析片段差
異,此法可取得不同處理間的具相異性片
段,回收這些片段作序列分析及相似性比
對,即可知道這些相異片段為哪些基因的
部分序列並推得受處理的檢體中基因層面
所受的影響。
在此研究中,我們針對這不同處理的7
個檢體以兩端各八個引子(共16 個引子),
進行排列組合進行片段放大工作,獲得64
種組合結果,再接者將放大反應得到的產
物以電泳膠分析後,選取具差異性意義之
片段,並切下、給予編號保留此片段,然
後對照以其引子對再次放大進行序列分
析,此階段共獲得126 個具處理間差異的
片段進行定序工作。
經過DNA 定序以及比對的結果,總
共獲得68 個在不同的處理組合中有不同
表現量的基因,所包含的基因種類有:代
謝,生長相關,肝臟再生,結構性蛋白質,
神經生成,凝血機制相關,訊息傳導,細
胞連結,原致癌基因,基因表現調控,發
炎反應,血液生成相關。
這些基因隨著不同的處理方法,呈現
出了不同的RNA 表現,可見其中應有與肝
臟術後再生有關的基因表現物。 | zh_TW |
dc.description.abstract | In this project, liver resection after a
period of ischemia in rats will be used as a
model to study the relation between ischemia
before hepatectomy and liver regeneration
after hepatectomy. And mRNA from the
samples will be extracted which stored in -70
℃ and double stranded cDNA will be
synthesized for amplified fragment length
polymorphism (AFLP) analysis.
To prepare an AFLP template, the
restriction fragments for amplifications are
generated by two different restriction
endonucleases. When used together, these
enzymes generate small cDNA fragments
that will be amplified well and are in the
optimal size range (<1Kb) for separation on
denaturing polyacrylamide gels. Following
heat inactivation of the restriction
endonucleases, the cDNA fragments are
ligated to adapters to generate template DNA
for amplification. PCR is performed in two
consecutive reactions.
In the first reaction, called
preamplification, cDNAs are amplified with
AFLP primers, each having one selective
nucleotide. The PCR products of the
preamplification reaction are diluted and
used as a template for the selective
amplification using two AFLP primers, each
containing three selective nucleotides. This
two-step amplification strategy results in
consistently cleaner and more reproducible fingerprints with the added benefit of
generating enough templates DNA for
thousands of AFLP reactions. Products from
the selective amplification are separated on a
denaturing polyacrylamide (sequencing) gel.
This technique would be taken
dissimilarity fragments among the samples
that could be reclaimed to sequence their
DNA sequences and similar comparing, so
we may understand how the genes expression
during these treatment. | en |
dc.format | application/pdf | zh_TW |
dc.format.extent | 54370 bytes | - |
dc.format.mimetype | application/pdf | - |
dc.language | zh-TW | zh_TW |
dc.language.iso | zh_TW | - |
dc.publisher | 臺北市:國立臺灣大學醫學院外科 | zh_TW |
dc.rights | 國立臺灣大學醫學院外科 | zh_TW |
dc.subject | 引子 | zh_TW |
dc.subject | 片段多型性分析 | zh_TW |
dc.subject | 肝臟 | zh_TW |
dc.subject | 缺血/灌流 | zh_TW |
dc.subject | AFLP | en |
dc.subject | liver | en |
dc.subject | ischemia/reperfusion | en |
dc.title | 利用AFLP技術探究缺氧對肝臟部份切除後再生之影響及基因表現之變化(3/3) | zh_TW |
dc.type | report | en |
dc.identifier.uri.fulltext | http://ntur.lib.ntu.edu.tw/bitstream/246246/24489/1/912314B002164.pdf | - |
dc.coverage | 計畫年度:91;起迄日期:2002-08-01/2003-07-31 | zh_TW |
item.cerifentitytype | Publications | - |
item.languageiso639-1 | zh_TW | - |
item.openairetype | report | - |
item.fulltext | with fulltext | - |
item.grantfulltext | open | - |
item.openairecristype | http://purl.org/coar/resource_type/c_93fc | - |
顯示於: | 醫學系
|