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題名 | 利用AcMNPV非寄主細胞之侷限性發展非裂解性桿狀病毒蛋白表現系統=Transduction of AcMNPV Nonpermissive Cells as a Non-lytic Baculovirus Expression System |
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作者姓名(中文) | 吳宗遠; 趙裕展; 李景欽; | 書刊名 | 臺灣昆蟲 |
卷期 | 24:1 2004.03[民93.03] |
頁次 | 頁15-26 |
分類號 | 369.74 |
關鍵詞 | 加州苜蓿夜蛾核多角體病毒; 非寄主細胞; 非裂解性桿狀病毒表現系統; AcMNPV; Nonpermissive cells; Non-lytic baculovirus expression system; |
語文 | 中文(Chinese) |
中文摘要 | 分析帶有加州苜蓿夜蛾核多角體病毒(Autographa californica multiple capsid nucleopolyhedrovirus,AcMNPV)早期啟動子IE1hr5、晚期啟動子VP39、最晚期啟動子P10及源自果蠅的hsp70啟動子驅動egfp螢光蛋白基因的重組AcMNPV病毒分別感染其寄主細胞SF21和非寄主細胞BmN4與SL2,顯示雖然BmN4和SL2可以抑制晚期基因的表現,但對早期基因如IE1和hsp70啟動子卻無明顯的抑制現象。因此BmN4和SL2細胞對AcMNPV之吸附性、進入及早期基因的表現可提供完整的支持。而源自果蠅的hsp70啟動子在AcMNPV感染SL2細胞後可持續的表現EGFP報導基因最少可維持8天之久,因此應可利用果蠅的hsp70啟動子和AcMNPV非寄主細胞SL2建立非裂解性桿狀病毒表現系統。 |
英文摘要 | Activities of viral and insect promoters constructed in an Autographa californica multiple capsid nucleopolyhedrovirus (AcMNPV) were tested in AcMNPV permissive-SF21AE cells and -nonpermissive BmN4 and SL2 cells. Recombinant AcMNPVs containing reporter genes (green fluorescence protein egfp gene) under the control of an early promoter (IE1hr5), a late promoter (VP39), a very late promoter (P10) of AcMNPV, and a heat shock promoter (hsp70), derived from Drosophila melanogaster, were constructed, respectively. In BmN4 and SL2, late and very late promoter activities were inhibited, but they expressed the early and hsp70 promoters. These results indicate that BmN4 and SL2 cells support the absorption, entry, and early gene expression of recombinant AcMNPVs. Interestingly, we found that the hsp70 promotercontaining recombinant AcMNPV expressed egfp at least 8 days after infection; this result suggests that SL2 cells can serve as host cells for recombinant AcMNPV in non-lytic baculovirus expression systems. |
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