Recombinant expression and reconstitution of multiprotein complexes by the USER cloning method in the insect cell-baculovirus expression system

被引：41

作者：

Zhang, Ziguo ^{[1
]}

Yang, Jing ^{[1
]}

Barford, David ^{[1
]}

机构：

[1] MRC, Mol Biol Lab, Francis Crick Ave, Cambridge CB2 0QH, England

来源：

METHODS | 2016年 / 95卷

关键词：

Baculovirus insect cell expression; MultiBac; Multi-protein complexes; USER ligation-free cloning; ANAPHASE-PROMOTING COMPLEX; URACIL-EXCISION; IN-VITRO; ARCHITECTURE; MECHANISM; APC/C; DNA; FRAGMENTS; BIOLOGY; VECTOR;

D O I：

10.1016/j.ymeth.2015.10.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The capacity to reconstitute complex biological processes in vitro is a crucial step in providing a quantitative understanding of these systems. It provides material for structural, biochemical and biophysical analyses and allows the testing of biological hypotheses and the introduction of chemical probes and tags for single molecule analysis. Reconstitution of these systems requires access to homogenous components, usually through their over-production in heterologous over-expression systems. Here we describe the application of the USER (Uracil-Specific Excision Reagent) ligation-free cloning method to assemble recombinant MultiBac transfer vectors for the generation of recombinant baculovirus suitable for the expression of multi-protein complexes in insect cells. Crown Copyright (C) 2015 Published by Elsevier Inc. All rights reserved.

引用

页码：13 / 25

页数：13

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