Grabherr R, Ernst W, Oker-Blom C, Jones I
University of Agriculture, Institute of Applied Microbiology, Muthgasse 18, A-1190, Vienna, Austria.
Trends Biotechnol. 2001 Jun;19(6):231-6. doi: 10.1016/s0167-7799(01)01610-9.
The ability to couple genotype to phenotype has proven to be of immense value in systems such as phage display and has allowed genes encoding novel functions to be selected directly from complex libraries. However, the complexity of many eukaryotic proteins places a severe constraint on successful display in Escherichia coli. This restriction could be resolved if a eukaryotic virus could be similarly engineered for display purposes. Preliminary data have suggested that the baculovirus Autographa californica, a multiple nuclear polyhedrosis virus (AcMNPV) is a candidate for eukaryotic virus display because the insertion of peptides into the native virus coat protein, or the expression of foreign proteins as coat protein fusions, results in incorporation of the sequence of interest onto the surface of virus particles. A variety of strategies are currently under investigation to develop further the display capabilities of AcMNPV and to improve the complexity of library that might be accommodated. Several expression vectors for different forms of surface display have been developed and, coupled with improved recombination strategies, represent progress towards a refined tool for use in functional genomics and in vitro protein evolution.
事实证明，在噬菌体展示等系统中，将基因型与表型联系起来的能力具有巨大价值，它使得编码新功能的基因能够直接从复杂文库中筛选出来。然而，许多真核生物蛋白质的复杂性对在大肠杆菌中成功展示构成了严重限制。如果真核病毒能够以类似方式进行工程改造用于展示目的，那么这一限制或许可以得到解决。初步数据表明，杆状病毒苜蓿银纹夜蛾多粒包埋型核型多角体病毒（AcMNPV）是真核病毒展示的一个候选对象，因为将肽插入天然病毒衣壳蛋白，或者将外源蛋白作为衣壳蛋白融合体表达，会导致目标序列整合到病毒颗粒表面。目前正在研究多种策略，以进一步拓展AcMNPV的展示能力，并提高其所能容纳文库的复杂性。已经开发出了几种用于不同形式表面展示的表达载体，再结合改进的重组策略，代表了在功能基因组学和体外蛋白质进化中使用的精细工具方面取得的进展。
