High-level expression of secreted glycoproteins in transformed lepidopteran insect cells using a novel expression vector

Farrell, Patrick J.; Lu, Maolong; Prevost, Jay; Brown, Chris; Behie, Leo A.; Iatrou, Kostas

doi:10.1002/(sici)1097-0290(19981220)60:6<656::aid-bit2>3.0.co;2-9

Cited by 90 publications

(86 citation statements)

References 32 publications

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“…The expression cassette pIE1/153A (23,45) was employed for expression of recombinant cystatin 1 in insect cells. Plasmid pIE1/153A.cyst was generated as follows.…”

Section: Methodsmentioning

confidence: 99%

A Virus Essential for Insect Host-Parasite Interactions Encodes Cystatins

Espagne¹,

Douris

Lalmanach

et al. 2005

J Virol

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“…The expression cassette pIE1/153A (23,45) was employed for expression of recombinant cystatin 1 in insect cells. Plasmid pIE1/153A.cyst was generated as follows.…”

Section: Methodsmentioning

confidence: 99%

A Virus Essential for Insect Host-Parasite Interactions Encodes Cystatins

Espagne¹,

Douris

Lalmanach

et al. 2005

J Virol

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“…PCR product was digested with NotI restriction endonuclease and was cloned in-frame into unique NotI site of pIE1/153A.jhe vector (18) and confirmed by sequencing. Culture of insect cells and procurement of samples containing recombinant protein from transfected insect cells are as described earlier (19). Briefly, High Five insect cells were seeded into six-well culture plates (35 mm diameter) at a density of 5 ϫ 10 5 cells/ml (2 ml/well) and transfected for 5 h with 0.5 ml of transfection solution containing 30 g/ml Lipofectin and 6 g/ml total plasmid DNA in basal IPL-41 medium.…”

Section: Tigeirdqmentioning

confidence: 99%

Functional Analysis of Cathepsin B-like Cysteine Proteases fromLeishmania donovani Complex

Somanna¹,

Mundodi²,

Gedamu

2002

Journal of Biological Chemistry

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Cathepsin B-like genes from Leishmania donovani and Leishmania chagasi have been isolated and characterized. It is a single gene, which is constitutively expressed in all the life cycle stages of the parasite. Studies using cathepsin B-specific inhibitor treatment suggested that cathepsin B does not seem to play a role in the promastigote stages of the parasite, however it aids in the parasite survival within the host macrophages. Antisense mRNA inhibition of cathepsin B gene also revealed that it plays an important role in the parasite survival within the host macrophages. Furthermore, for the first time, we have shown that Leishmania whole cell lysates as well as the recombinant cathepsin B protein cleaved human recombinant latent transforming growth factor (TGF)-␤1 into a mature peptide releasing the latency associated protein, in a cell-free incubation system. Mink lung epithelial cell growth inhibition assay revealed that the cleaved TGF-␤1 was biologically active, suggesting that Leishmania cathepsin B can cleave latent TGF-␤1 into mature and active form. These results suggest that cathepsin B plays an important role in Leishmania survival within the host macrophages by activating latent TGF-␤1.

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“…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. 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.…”

mentioning

confidence: 99%

“…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.…”

mentioning

confidence: 99%