Production of Multiprotein Complexes Using the Baculovirus Expression System: Homology-Based and Restriction-Free Cloning Strategies for Construct Design.

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

https://www.ncbi.nlm.nih.gov/pubmed/33301110

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