改造鳞翅目昆虫细胞以改善蛋白质加工和表达

Transforming Lepidopteran Insect Cells for Improved Protein Processing and Expression.

作者信息

Harrison Robert L, Jarvis Donald L

机构信息

Invasive Insect Biocontrol & Behavior Laboratory, USDA, ARS, BARC, Building 007, Room 301, BARC-W, 10300 Baltimore Avenue, Beltsville, MD, 20705, USA.

Department of Molecular Biology, University of Wyoming, Laramie, WY, USA.

出版信息

Methods Mol Biol. 2016;1350:359-79. doi: 10.1007/978-1-4939-3043-2_18.

DOI:10.1007/978-1-4939-3043-2_18

PMID:26820868

Abstract

The lepidopteran insect cells used with the baculovirus expression vector system (BEVS) are capable of synthesizing and accurately processing foreign proteins. However, proteins expressed in baculovirus-infected cells often fail to be completely processed, or are not processed in a manner that meets a researcher's needs. This chapter discusses a metabolic engineering approach that addresses this problem. Basically, this approach involves the addition of new or enhancement of existing protein processing functions in established lepidopteran insect cell lines. In addition to improvements in protein processing, this approach has also been used to improve protein expression levels obtained with the BEVS. Methods for engineering cell lines and assessing their properties as improved hosts for the BEVS are detailed. Examples of lepidopteran insect cell lines engineered for improved protein N-glycosylation, folding/trafficking, and expression are described in detail.

摘要

与杆状病毒表达载体系统（BEVS）一起使用的鳞翅目昆虫细胞能够合成并准确加工外源蛋白。然而，在杆状病毒感染细胞中表达的蛋白常常无法被完全加工，或者加工方式不能满足研究人员的需求。本章讨论一种解决该问题的代谢工程方法。基本上，该方法涉及在已建立的鳞翅目昆虫细胞系中添加新的或增强现有的蛋白加工功能。除了改善蛋白加工外，该方法还被用于提高通过BEVS获得的蛋白表达水平。详细介绍了工程化细胞系的方法以及评估它们作为BEVS改良宿主的特性。详细描述了为改善蛋白N-糖基化、折叠/运输和表达而进行工程改造的鳞翅目昆虫细胞系实例。