線形噬菌體在組裝時所牽涉的蛋白質-蛋白質及蛋白質-DNA間交互作用之分析

Abstract

線形噬菌體是一種微小且高度進化的寄生蟲，具有單股環狀DNA基因組，它在繁衍及散佈時不會殺死宿主細胞。其組裝與釋出細胞膜是相伴進行，且在此過程中，可能有牽涉到一連串蛋白質-蛋白質與蛋白質-DNA的交互作用。在之前研究發現，去掉線形噬菌體包裝信息所造成的傷害，可以被g7p、g9p、及g1p的突變給補償。代表這些蛋白質可能與組裝起始相關。除此之外，thioredoxin對於繁衍過程也是必須的，且可能會與g1p作用來啟動組裝的機制。這篇報告利用膠體遷移分析來檢視前人提出有關包裝信息-蛋白質間可能之交互作用。我們同時也藉由各種分析檢測這些相關蛋白質之間可能之交互作用。這些結果對於線形噬菌體在結構及組裝過程提供了相關訊息。

Filamentous phages are small , highly evolved parasites that can reproduce and disseminate without killing their host . The genome of filamentous phage is single stranded circular DNA . The process by which filamentous phage are concomitantly assembled and secreted across the cell membranes is likely to involve a series of protein – protein and protein – DNA interactions .The previous findings that deletion of the filamentous phage packaging signal can be compensated for by mutations in g7p , g9p , and g1p , strongly suggest a role for these proteins in the initiation of assembly . In addition , thioredoxin is required for efficient filamentous phage production and may interact to g1p to drive an engine for filamentous phage assembly . This report tests the gel shift assay for examining previous proposal about protein- packaging signal interaction . We also investigate the possible interactions between these proteins by various assays. These results would show information on the structure and assembly process of filamentous phage .