| 摘要 |
#CMT# #/CMT# A recombinant baculovirus resulting from insertion of expression cassettes, permitting protein expression in a host cell, into a baculovirus, comprising a first expression cassette comprising a primary promoter and a second expression cassette comprising a second promoter, in which the promoters being other than an endogenous baculovirus/host cell promoter and recognized by the RNA polymerase II of the host cell and the second promoter being different from the first promoter, is new. #CMT# : #/CMT# An independent claim is included for a host cell infected by the recombinant baculovirus. #CMT#USE : #/CMT# The baculovirus recombinant/host cell is useful for the production of glycoproteins (claimed) and for the production of protein complex. #CMT#ADVANTAGE : #/CMT# The production of protein by the baculovirus recombinant/host cell is simple, fast and cost effective and the process exhibits high yield. #CMT#BIOTECHNOLOGY : #/CMT# Preparation (Disclosed): Preparation of recombinant baculovirus comprises inserting glycosyltransferases genes in the baculovirus genome; cloning the target genes in the bacterial plasmid vector to obtain an expression cassette; inserting the expression cassette in viral genome by recombination; producing an intermediary baculovirus possessing a phenotype *(OB+); identifying the recombinant baculovirus based on the phenotype (OB+). Preferred Components: The recombinant baculovirus comprises a third expression cassette comprising a third promoter i.e. different from first and second promoters and is other than an endogenous baculovirus/host cell promoter and recognized by the RNA polymerase II of the host cell; additional expression cassettes, where each of the cassettes comprises a promoter other than an endogenous baculovirus/host cell promoter and recognized by the RNA polymerase II of the host cell and the promoters being different from each other; and expression cassettes comprising very late baculovirus promoters, preferably 3 expression cassettes allowing the three different glycosyltransferase expressions, where the promoters are different from each other. The expression cassettes are positioned in a region i.e. different from viral genome containing genes that are nonessential for the replication of the virus in cellular culture. #CMT#EXAMPLE : #/CMT# Sf9 cell line (ATCC CRL-1711) derived from Spodoptera frugiperdawas used to construct the recombinant baculovirus. The Sf9 cell was cultured at 28[deg]C in TCIOO medium. The Sf9 cell was infected with baculovirus ( Autographa californicamultiple nuclear polyhedrosis virus (AcMNPV)). The baculovirus was constructed by inserting the Sse8387I and Bsu36I sites. |
