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Effect of Temporal Expression of Integral Membrane Proteins by Baculovirus Expression Vector System

机译:杆状病毒表达载体系统整体膜蛋白的时间表达的影响

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摘要

Integral membrane proteins (IMPs) are popular target for drugs, but their resolved structures have been overlooked when compared with cytosolic proteins. The main reason is that IMPs usually need intensive post-translational modifications and they are bound to membranes, which increase the complexity of purifying or crystalizing them. Although different expression systems are used to express IMPs, baculovirus is considered one of the most successful expression systems for those proteins. Despite that, there are always unknown discrepancies in the level of IMPs expression in the baculovirus expression system. Retrospective studies have shown that expression of an immunoglobulin (anti-Chymase mouse monoclonal IgG1) driven by vp39 promoter was more efficient compared to its expression under polyhedrin (polh) promoter; however, this conclusion was not tested on different IMPs to generalize such a conclusion. In this study, the expression of eight different IMPs has been compared under vp39 and polh promoters of Autographa californica nucleopolyhedrovirus. Although different IMPs have shown different patterns of expression, the expression driven by vp39 promoter was found to be generally more efficient than the polh promoter.
机译:整体膜蛋白(Imps)是药物的普遍靶标,但与细胞溶质蛋白相比,它们的分离结构被忽略了。主要原因是IMBS通常需要密集的翻译后修饰,并且它们与膜结合,这增加了净化或晶体化的复杂性。虽然不同的表达系统用于表达IMPS,但是杆状病毒被认为是这些蛋白质最成功的表达系统之一。尽管如此,在杆状病毒表达系统中存在百分比表达的差异存在差异。回顾性研究表明,与其在多溴苯胺(POLH)启动子下的表达相比,由VP39启动子驱动的免疫球蛋白(抗冠蛋白小鼠单克隆IgG1)的表达更有效;然而,这一结论未在不同的IMP上进行测试,以概括这样的结论。在这项研究中,在VP39和Polh启动子下比较了八种不同的Imps的表达,并在纯粹的术语中的核心核细胞核血症的VP39和Polh启动子。虽然不同的IMBs表现出不同的表达模式,但发现由VP39启动子驱动的表达通常比POLH启动子更有效。

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