Production of Multiprotein Complexes Using the Baculovirus Expression System: Homology-Based and Restriction-Free Cloning Strategies for Construct Design.
Fiche publication
Date publication
janvier 2021
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Mr ESSABRI Karim, Dr POTERSZMAN Arnaud, Mme ROSSOLILLO Paola
Tous les auteurs :
Rossolillo P, Kolesnikova O, Essabri K, Zamorano GR, Poterszman A
Lien Pubmed
Résumé
Most cellular processes are mediated by multi-subunit protein complexes which have attracted major interest in both academia and industry. Recombinant production of such entities in quantity and quality sufficient for functional and structural investigations may be extremely challenging and necessitate specific technologies. The baculovirus expression vector system is widely used for the production of eukaryotic multiprotein complexes, and a variety of strategies are available to assemble transfer vectors for the generation of recombinant baculoviruses. Here we detail applications of homology-based cloning techniques for one-step construction of dual promoter baculovirus transfer plasmids and of restriction-free (RF) cloning for the modification of existing constructs.
Mots clés
Baculovirus insect cell expression, Homology-based cloning, Multiprotein complex, Sequence and ligation–independent cloning (SLIC ), restriction-free (RF ) cloning
Référence
Methods Mol Biol. 2021 ;2247:17-38