Recombinant expression and reconstitution of multiprotein complexes by the USER cloning method in the insect cell-baculovirus expression system.
Methods
; 95: 13-25, 2016 Feb 15.
Article
de En
| MEDLINE
| ID: mdl-26454197
ABSTRACT
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.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Plasmides
/
Baculoviridae
/
Clonage moléculaire
/
Transgènes
/
Complexes multiprotéiques
/
Vecteurs génétiques
Limites:
Animals
Langue:
En
Journal:
Methods
Sujet du journal:
BIOQUIMICA
Année:
2016
Type de document:
Article
Pays d'affiliation:
Royaume-Uni