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胞裂性與非胞裂性桿狀病毒系統表現膜蛋白之比較與研究

胞裂性與非胞裂性桿狀病毒系統表現膜蛋白之比較與研究.

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胞裂性與非胞裂性桿狀病毒系統表現膜蛋白之比較與研究

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  1. 胞裂性與非胞裂性桿狀病毒系統表現膜蛋白之比較與研究 • 為探討非胞裂性桿狀病毒表現系統之優點及可行性，在本研究中使用5’-BrdU突變重組病毒vABhcmEpL，並挑選具有最明顯非胞裂性之重組病毒株C4，與 polh (AcMNPV多角體)起動子活性最強的重組病毒株PN24，以探討非胞裂性桿狀病毒表現系統與胞裂性桿狀病毒表現系統。vABhcmEpL係AcMNPV之重組病毒，含有由CMV minimal 起動子(promoter)起動、經hr提供加強子(enhancer)活性(hCMVm)之EGFP基因，以及由polh起動子起動之螢光酵素(luciferase)基因。此二基因在此扮演報導基因的角色。將阿拉伯芥(Arabidopsis thaliana)之鈉離子通道蛋白質CHL1之N端穿膜區域(transmembrane domains)與圓盤擬珊瑚海葵(Discosoma sp.)紅螢光蛋白質DsRed接合，得一融合蛋白質CHL1-DsRed。再將vABhcmEpL, C4, PN24等病毒株再度重組，從每一個病毒株中各構築出二株重組病毒，分別含有由hCMVm或polh起動子起動之CHL1-DsRed蛋白質。這些重組病毒感染Sf21細胞後，以顯微鏡觀察並用螢光光度計測量螢光強度，觀測 hCMVm與polh起動子的效果，並且，利用西方點墨法分析細胞殘渣與上清液。試驗結果顯示以膜蛋白而言，早期起動子hCMVm之生產量比晚期起動子polh高；而非胞裂性桿狀病毒表現系統比傳統胞裂性桿狀病毒表現系統之生產量高。

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