YBac: A novel and simple baculovirus system for heterologous protein production in Spodoptera frugiperda 9 (Sf9) cells.
baculovirus
gene deletion
heterologous protein expression
homologous recombination
insect cells
Journal
Biotechnology journal
ISSN: 1860-7314
Titre abrégé: Biotechnol J
Pays: Germany
ID NLM: 101265833
Informations de publication
Date de publication:
Jul 2023
Jul 2023
Historique:
revised:
06
03
2023
received:
02
11
2022
accepted:
29
03
2023
medline:
10
7
2023
pubmed:
6
4
2023
entrez:
5
4
2023
Statut:
ppublish
Résumé
The baculovirus expression vector system (BEVS) has been widely used for heterologous protein expression due to its powerful functionality and easy access to commercial expression vectors. Currently, most laboratories prefer two strategies for protein production using BEVS. One is recombinant bacmid based on transposition in Escherichia coli (e.g., Bac-to-Bac), and another is homologous recombination in insect cells (e.g., flashBac). In this manuscript, a rapid and simple YBac system was established. This novel system uses an Ac99KO bacmid as a virus vector and co-transfected into Spodoptera frugiperda 9 (Sf9) cells with a donor plasmid capable of recombination into Ac42 loci that carry the genes of interest (GOIs) along with the complete Ac99 fragment. Based on the intracellular homologous recombination system, the production of foreign proteins was achieved by the complementation of the Ac99 gene together with the insertion of GOIs. In this study, the human thyroid peroxidase (hTPO) and porcine epidemic diarrhea virus-like particles (PEDV VLPs) were successfully expressed using the YBac system. The entire process was shortened to 10 days, and the components involved in the system could be easily prepared in the laboratory, suggesting that the YBac system may have great potential in the production of heterologous proteins.
Identifiants
pubmed: 37016491
doi: 10.1002/biot.202200558
doi:
Substances chimiques
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2200558Informations de copyright
© 2023 Wiley-VCH GmbH.
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