Stably Transformed Insect Cell Lines: Tools for Expression of Secreted and Membrane‐anchored Proteins and High‐throughput Screening Platforms for Drug and Insecticide Discovery

Production of Japanese encephalitis virus-like particles in insect cells

“…The activity of promoters used for insect cell expression systems can be enhanced by certain cis-or trans-acting elements derived from baculoviruses. 19 Recently, an expression vector containing the Bombyx mori cytoplasmic actin promoter, from which foreign gene expression is stimulated with the B. mori NPV (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer, has been developed for high-level expression of recombinant proteins in lepidopteran insect cells. [16][17][18] Use of the IE-1 transactivator and the HR3 enhancer with the actin promoter has resulted in a more than 1000-fold increase in the stimulation of foreign gene expression through the actin promoter.…”

“…[16][17][18] Use of the IE-1 transactivator and the HR3 enhancer with the actin promoter has resulted in a more than 1000-fold increase in the stimulation of foreign gene expression through the actin promoter. 19,27 We used plasmid vectors that contained the B. mori actin promoter downstream from the BmNPV IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression, together with either a blasticidin or a neomycin resistance gene for use as a selectable marker (Fig. 1).…”

“…9 Stably transformed insect cells allow the constitutive production of recombinant proteins and can be employed as attractive alternative platforms to the baculovirus-insect cell system. 13,[16][17][18][19][20] They are particularly useful for the production of secreted complex proteins, because the protein synthesis and processing machinery of the host insect cell is not compromised by baculoviral infection. Recently, we investigated the production of JE VLPs in stably transformed lepidopteran insect cells.…”

Abnormal development in larvae of Sesamia nonagrioides (Lepidoptera: Noctuidae) resulting from baculovirus-mediated overexpression of a JHE-related gene (SnJHER)

“…PCR was performed using the JHERHetEF/JHERHetER primer pair (Table 1) to amplify 1730 bp of the SnJHER cDNA and introduce fl anking NotI recognition sequences and the consensus Kozak motif (GCCACC, light-grey shaded) just before the ATG codon (dark-grey shaded and in bold) ( Table 1). The amplifi ed PCR product was digested using NotI (New England Biolabs) and ligated into the NotI position of the pEIA-N-Flag expression vector (Douris et al, 2006) to generate the pEIA-N-Flag/SnJHER expression plasmid.…”

Lepidopteran cells, an alternative for the production of recombinant antibodies?

“…However this system presents a significant advantage, that secreted proteins are processed and secreted more efficiently. 71 In 2000, Guttieri et al 72,73 have used this new technology to produce antibodies. Sf 9 cells were transfected with 3 independent constructs, each containing one expression cassette for H, L and the neomycin resistance gene.…”