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研究生: 林淑珍
Soo-Chen Lin
論文名稱: Hz-1病毒極早期基因的分子選殖
Molecular Cloning of Very Early Gene of Hz-1 Virus
指導教授: 李銘亮
Li, Ming-Liang
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
畢業學年度: 83
語文別: 中文
論文頁數: 56
中文關鍵詞: 桿狀病毒;極早期基因;分子選殖;Hz-1病毒
英文關鍵詞: Baculovirus;Very early gene;Molecular cloning;Hz-1 virus
論文種類: 學術論文
相關次數: 點閱:232下載:0
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  • Hz-1 病毒是Baculovirus 的一種。由於它是所有動植物裡面極少數能在不
    同細胞株中進行明顯的急性及持續感染 ( productive and persistent
    infection ) 的病毒。因此,成為研究病毒急性及持續感染機制的良好模
    式。為瞭解病毒急性及持續感染的分子轉換機制,有必要比較急性特異性
    及持續感染相關性之起動子,以便知道其控制機制。由於持續感染相關性
    起動子已被發現,若能找到急性感染特異性起動子,則將極有助於此轉換
    機制的研究。Hz-1 病毒之絕大部分極早期基因皆位於 HindIII-A、D、I
    三片段上,但由於其可能對細菌有毒,以前曾有學者用質體,cosmid,及
    phage 皆無法選殖成功。於是本實驗改用 low copy number 的 pWSK-29
    質體,才選殖到 HindIII-A、D兩片段。Southern analysis 的結果確定質
    體所插入之 DNA 確實分別為 16.1 kb 的 HindIII-A ( HA ) 及 13.4 kb
    的HindIII-D ( HD ) 片段。此二質體的限制酵素圖譜已被完成,在 HD 片
    段上的所有轉錄子 ( transcripts ) 及其方向也完整的經過解析。HD 片
    段區域內含有 1.35 kb、1.52 kb、1.78 kb、2.33 kb 及 2.91 kb 的早期
    轉錄子及 4.60 kb、4.81 kb、6.05 kb 及 7.33 kb 的晚期轉錄子。HD 片
    段並且含有一個極早期基因,此基因轉錄了一個 5.54 kb 的轉錄子。這個
    極早期基因經進一步分析發現位於 XhoI-H 片段到 XhoI-W 片段的區域,
    其轉錄方向已確定。而 5.54 kb 的轉錄子上游含有 TATA 及 CAGT 之區域
    則可能是起動子。此為 Hz-1病毒之第一個被發現的極早期基因及起動子

    Hz-1 virus is a non-occluded baculovirus. It is one of the
    few animal and plant viruses which can induce both productive and
    persistent viral infections in the culture cells. In order to
    understand the molecular switches of productive and persistent
    viral infections, promoters specifically associated to these two
    phases should be compared. Since the persistent-associated gene
    is driven by an very early type promoter, the cloning of
    productive-specific very early promoters is needed. According to
    previous results, most of the very early type genes of Hz-1 virus
    are located on HindIII-A, D, and I fragments. For some unknown
    reasons, these fragments are not possible to be cloned by
    conventional plasmids and cosmids.However in my study, HindIII-A
    and D fragments are successfully cloned by using a low copy
    number plasmid, pWSK-29. Southern analysis confirms that the two
    viral DNA fragments, HindIII -A of 16.1kb and HindIII-D of 13.4
    kb, have been inserted into pWSK-29 plasmids. Restriction maps of
    these two contructed plasmids are produced and all the
    transcripts and their directions on the HD fragment have been
    identified. Hind III-D fragment contains early and late
    transcripts. A very early gene, which transcripts a 5.54kb RNA,
    was also found to be contained in this fragment. This very ealy
    gene is located between XhoI-H and XhoI-W fragments of viral
    genome. Furthermore, the promotor may locate at the TATA and CAGT
    areas upstream of 5.54 kb transcript. These are the first
    productive-infection associated very early gene and promotor
    being reported in Hz-1 virus.
    Hz-1 virus is a non-occluded baculovirus. It is one of the

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